Research Article |
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Corresponding author: Nisarat Tungpairojwong ( knisar@kku.ac.th ) Academic editor: Pavel Stoev
© 2026 Chalintorn Wilaisorn, Ruttapon Srisonchai, Wibhu Kutanan, Nisarat Tungpairojwong.
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.
Citation:
Wilaisorn C, Srisonchai R, Kutanan W, Tungpairojwong N (2026) Two new species and two new country records of the giant pill-millipede genus Zephronia Gray, 1832 from Thailand, based on integrative taxonomy (Diplopoda, Sphaerotheriida, Zephroniidae). ZooKeys 1294: 1-59. https://doi.org/10.3897/zookeys.1294.205518
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An integrative taxonomic study combining morphological and molecular data revealed two new species of giant pill-millipedes of the genus Zephronia Gray, 1832: Zephronia tamarindina Wilaisorn & Srisonchai, sp. nov. and Zephronia triforma Wilaisorn & Srisonchai, sp. nov., both endemic to provinces in the upper central region of Thailand. In addition, Zephronia dawydoffi Attems, 1953 and Zephronia laotica Wesener, 2019 are recorded from Thailand for the first time. The new species exhibit distinctive colouration patterns, and scanning electron microscopy revealed unique morphological characters for each taxon. Analysis of the 658 bp DNA barcoding region of the mitochondrial COI gene showed interspecific genetic distances ranging from 8.68–21.92% for Z. dawydoffi, 12.79–22.98% for Z. laotica, 10.81–22.07% for Z. tamarindina sp. nov., and 10.81–22.83% for Z. triforma sp. nov., in comparison with closely related species from Thailand, Vietnam, Laos, and Cambodia. Phylogenetic analyses placed all four species within the Zephronia s.s. clade, characterised by the Tömösváry’s organ located outside the antennal groove. However, relationships among other clades remained unresolved due to interspersion with species of Cryxus Leach, 1814, Prionobelum Verhoeff, 1924, and Sphaerobelum Verhoeff, 1924. Zephronia was recovered as non-monophyletic, indicating that the current generic limits within Zephroniinae require reassessment using multi-locus data. These results expand our understanding of giant pill-millipede diversity and highlight the upper central Thai region and Southeast Asia more broadly as a significant biodiversity hotspot.
Biodiversity, DNA barcoding, Myriapoda, Southeast Asia, taxonomy
The order Sphaerotheriida with their family Zephroniidae are notable for their heavily sclerotised exoskeleton and remarkable ability to roll into a ball-like shape when threatened (
Thailand, situated within the Indo-Burma biodiversity hotspot, harbours a remarkable diversity of millipedes (Diplopoda), with approximately 287 recorded species, a continuously growing number as new discoveries are made (
Recent targeted surveys have discovered many new species of the genus Zephronia. Since 2021, the interest in Thai giant pill-millipedes has surged, leading to the recognition of up to 14 Zephronia species, viz., Z. chantaburiensis Srisonchai & Wesener, 2024, Z. chrysomallos Bhansali & Wesener, 2022, Z. enghoffi Srisonchai & Likhitrakarn, 2021, Z. erawani Bhansali & Wesener, 2022, Z. golovatchi Srisonchai & Likhitrakarn, 2021, Z. lannaensis Likhitrakarn & Golovatch, 2021, Z. macula Srisonchai & Wesener, 2024, Z. ovalis Gray, 1832, Z. panhai Srisonchai & Likhitrakarn, 2021, Z. phrain Likhitrakarn & Golovatch, 2021, Z. siamensis Hirst, 1907, Z. sukhothaiensis Srimongkol & Srisonchai, 2026, Z. tratensis Srimongkol & Srisonchai, 2026, and Z. viridisoma Rosenmejer & Wesener, 2021. This growing list underscores the remarkable morphological diversity of Zephronia in Thailand. However, as with other genera in the Zephroniidae, the taxonomic research on Zephronia remains challenging due to limited genetic data and unclear evolutionary relationships (
DNA barcoding, based on a fragment of the mitochondrial cytochrome c oxidase subunit I (COI) gene, has become a powerful molecular tool for millipede species identification. This method enables the rapid and reliable detection of genetic divergence, even among morphologically similar taxa, which is particularly valuable for groups with cryptic species or subtle diagnostic traits. When combined with morphological data, DNA barcoding provides a robust framework for resolving taxonomic uncertainties, verifying species boundaries, and uncovering hidden diversity. Such integrative approaches have proven especially effective for species delimitation for the giant pill-millipede genus Zephronia (
Continuous field surveys and taxonomic workshops have resulted in the accumulation of extensive material over time, from which the specimens examined in this study were obtained. Examination of four morphologically distinct groups of giant pill-millipedes collected during field surveys in 2022–2024 revealed both previously undescribed species and new country records. These findings highlight the rich but still incompletely documented diversity of Zephronia in Thailand, particularly in underexplored habitats where endemism is likely high. In this study, we formally describe two new Zephronia species and report the first records of Z. dawydoffi Attems, 1953 and Z. laotica Wesener, 2019 in the country. To ensure accurate identification and resolve potential taxonomic ambiguities, we employed an integrative taxonomic approach, combining detailed morphological examination with molecular analysis based on mitochondrial COI sequences.
The sampling was conducted across the upper central and northeastern regions of Thailand, as well as adjacent areas between 2022 and 2024. All specimens were collected by hand, searching at ground level, beneath leaf litter, and within decomposed logs across a variety of habitats, including agricultural fields, plantation forests, and limestone mountains. Live specimens were photographed in situ using a Canon 90D digital camera equipped with a Canon EF 100 mm f/2.8 Macro USM lens. Specimens were euthanised in accordance following the guidelines of the
The map was generated using a background image from Simplemappr (
A total of 290 specimens were examined under a stereomicroscope. Species identification was based on morphological characters and comparisons with descriptions from the previous literature, as well as with type specimens of other Thai congeners. Morphological terminology and descriptive formats followed those used by
Type specimens, including holotypes and paratypes, were deposited in the following institutions: Chulalongkorn University Museum of Zoology, Thailand (
Because the present study focuses on the description and redescription of four species rather than a comprehensive revision of the genus Zephronia, no general description of the genus is provided. Instead, each species is described separately in full to facilitate direct comparisons and to ensure clarity in species delimitations. Such a format is commonly adopted in taxonomic works dealing with a limited number of new species, particularly when the primary objective is species diagnoses and identification rather than a generic revision.
3it 3-combed inner tooth of mandible,
bu bursa of vulva,
co condylus of mandible,
Cp central pad of gnathochilarium,
cr-t crenulated teeth on telopods,
ct central tooth,
cx coxa,
et external tooth of epipharynx,
Et external tooth of mandibular gnathal lobe,
fe femur,
Ia inner area of mandible,
il incisura lateralis of the head,
ip inner palpi of gnathochilarium,
ll lamellae lingulales of gnathochilarium,
me mentum of gnathochilarium,
ml membranous lobe,
mp molar plate of mandible,
Mp membranous process
op operculum of vulva,
pl pectinate lamellae of mandible,
po postfemur,
pre prefemur,
rdg femoral ridge,
rsp row of spines,
sc sensory cone,
scl-s sclerotised spots,
st stipites of gnathochilarium,
st-pl stigmatic plate,
syn-cx syncoxite,
ta tarsus,
ti tibia.
The specimens were examined and dissected under a stereo microscope. Morphological structures were dried in the desiccator for 24 hours, mounted on aluminium stubs, and coated with gold. Samples were then photographed under a high vacuum in a JEOL JSM-5410 LV scanning electron microscope at the Faculty of Science, Khon Kaen University. All figures were subsequently assembled and refined in Adobe Photoshop 2025.
The genomic DNA was extracted from 27 specimens, comprising four and eight specimens of the two new species, ten specimens of Z. dawydoffi, and foive specimens of Z. laotica. Extractions were performed from the leg tissue using proteinase K digestion and following the manufacturer’s instructions of Vivantis GF-1 Nucleic Acid Extraction Kits. PCR amplification of the 658 bp barcoding region of the mitochondrial cytochrome c oxidase subunit I (COI) gene using the universal primers — LCO-1490 (5’-GGT CAA CAA ATC ATA AAG ATA TTG G-3’) and HCO-2198 (5’-TAA ACT TCA GGG TGA CCA AAA AAT CA-3’) (
A total of 124 sequences were analysed, including 27 sequences generated in this study and 97 sequences retrieved from public databases representing all recorded Zephronia species. Outgroups included Arthrosphaera brandtii (Humbert, 1865), Cryxus ovalis (Linnaeus, 1758), Epicyliosoma sp., Glomeris marginata (Villers, 1789), Prionobelum spp., Sphaerobelum spp., and Sphaeromimus splendidus Wesener & Sierwald, 2005. The newly generated sequences were then deposited in GenBank (Table
| Species | COI accession number | Voucher code | Locality | Reference | |
|---|---|---|---|---|---|
| Ingroups | |||||
| 1 | Zephronia chantaburiensis Srisonchai & Wesener, 2024 | PP754582 |
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Thailand, Chantaburi Province, Tha Mai District, Wat Khao Sukim | |
| 2 | Zephronia chantaburiensis Srisonchai & Wesener, 2024 | PP754583 |
|
Thailand, Chantaburi Province, Tha Mai District, Wat Khao Sukim | |
| 3 | Zephronia chrysomallos Bhansali & Wesener, 2022 | OM509649 | ZFMK MYR8826 | Thailand, Kanchanaburi Province, Sai Yok District, Sai Yok Noi Waterfall | |
| 4 | Zephronia dawydoffi Attems, 1953 | MK330971 | ZFMK Myr4504 | N/A | |
| 5 | Zephronia dawydoffi Attems, 1953 | PX737475 | MZKKU (BRM06) | Thailand, Buri Ram Province, Khu Mueang District, Dong Kheng rubber plantation | This study |
| 6 | Zephronia dawydoffi Attems, 1953 | PX737476 | MZKKU (BRM10) | Thailand, Buri Ram Province, Khu Mueang District, Dong Kheng rubber plantation | This study |
| 7 | Zephronia dawydoffi Attems, 1953 | PX737477 | MZKKU (BRM17) | Thailand, Buri Ram Province, Khu Mueang District, Non Somboon Monastery | This study |
| 8 | Zephronia dawydoffi Attems, 1953 | PX737478 | MZKKU (BRM24) | Thailand, Buri Ram Province, Khu Mueang District, Non Somboon Monastery | This study |
| 9 | Zephronia dawydoffi Attems, 1953 | PX737479 | MZKKU (DBR01) | Thailand, Buri Ram Province, Mueang District, Khao Kradong | This study |
| 10 | Zephronia dawydoffi Attems, 1953 | PX737480 | MZKKU (DBR02) | Thailand, Buri Ram Province, Mueang District, Khao Kradong | This study |
| 11 | Zephronia dawydoffi Attems, 1953 | PX737481 | MZKKU (DBR04) | Thailand, Buri Ram Province, Mueang District, Ban Khok Ta Sing Waterfall | This study |
| 12 | Zephronia dawydoffi Attems, 1953 | PX737482 | MZKKU (DBR06) | Thailand, Buri Ram Province, Mueang District, Ban Khok Ta Sing Waterfall | This study |
| 13 | Zephronia dawydoffi Attems, 1953 | PX737483 | MZKKU (KKN12) | Thailand, Khon Kaen Province, Mueang District, Thung Setthi Temple | This study |
| 14 | Zephronia dawydoffi Attems, 1953 | PX737484 | MZKKU (KKN20) | Thailand, Khon Kaen Province, Mueang District, Thung Setthi Temple | This study |
| 15 | Zephronia erawani Bhansali & Wesener, 2022 | OM509650 |
|
Thailand, Kanchanaburi Province, Si Sawat District, 50 km W of Kanchanaburi, Erawan Waterfall | |
| 16 | Zephronia golovatchi Srisonchai, Sutcharit & Likhitrakarn, 2021 | OM509646 | ZFMK MYR6262 | Thailand, Nakhon Ratchasima Province, Pak Chong District | |
| 17 | Zephronia golovatchi Srisonchai, Sutcharit & Likhitrakarn, 2021 | OM509647 | ZFMK K53 | Thailand, Nakhon Nayok Province, Khao Yai National Park | |
| 18 | Zephronia hui Liu & Wesener, 2022 | OP339790 | SCAU YGM03 | China, Guizhou, Tongren City, Jiangkou County, Taiping Town, Yamugou Scenic Area | |
| 19 | Zephronia lannaensis Likhitrakarn & Golovatch, 2021 | OM509629 | ZFMK MYR3498 | Thailand, Chiangmai Province, Mueang District, Doi Suthep | |
| 20 | Zephronia lannaensis Likhitrakarn & Golovatch, 2021 | OM509630 | ZFMK MYR3501 | Thailand, Chiangmai Province, Mae Rim District, Traidhos School Campus | |
| 21 | Zephronia lannaensis Likhitrakarn & Golovatch, 2021 | OM509631 | ZFMK MYR4911 | Thailand, Chiangmai Province, Mae Rim District, Mae Sa Valley | |
| 22 | Zephronia lannaensis Likhitrakarn & Golovatch, 2021 | OM509632 |
|
Thailand, Chiangmai Province, Mueang District, Doi Suthep, Me Sa Waterfall | |
| 23 | Zephronia lannaensis Likhitrakarn & Golovatch, 2021 | OM509633 | MHNG 3B | Thailand, Chiangmai Province, Mueang District, Doi Suthep | |
| 24 | Zephronia laotica Wesener, 2019 | MK330977 | ZFMK Myr3502 | Laos, Champasak Province, east of Mekong, Garden of Erawan Riverside Hotel | |
| 25 | Zephronia laotica Wesener, 2019 | PX737485 | MZKKU (SSK01) | Thailand, Si Sa Ket Province, Khun Han District, Mae Lamoon Durian plantation | This study |
| 26 | Zephronia laotica Wesener, 2019 | PX737486 | MZKKU (SSK03) | Thailand, Si Sa Ket Province, Khun Han District, Mae Lamoon Durian plantation | This study |
| 27 | Zephronia laotica Wesener, 2019 | PX737487 | MZKKU (SSK14) | Thailand, Si Sa Ket Province, Khun Han District, Kantrom Tai Temple | This study |
| 28 | Zephronia laotica Wesener, 2019 | PX737488 | MZKKU (SSK18) | Thailand, Si Sa Ket Province, Khun Han District, Kantrom Tai Temple | This study |
| 29 | Zephronia laotica Wesener, 2019 | PX737489 | MZKKU (SSK25) | Thailand, Si Sa Ket Province, Khun Han District, Ta Nuan Rubber plantation | This study |
| 30 | Zephronia macula Srisonchai & Wesener, 2024 | PP754589 |
|
Thailand, Sra Kaeo Province, Mueang Sra Kaeo District, Wat Tham Khao Maka | |
| 31 | Zephronia macula Srisonchai & Wesener, 2024 | PP754590 |
|
Thailand, Sra Kaeo Province, Mueang Sra Kaeo District, Wat Tham Khao Maka | |
| 32 | Zephronia macula Srisonchai & Wesener, 2024 | PP754585 |
|
Thailand, Chantaburi Province, Khlung District, Thaeo Khlong Khlung Monastery | |
| 33 | Zephronia macula Srisonchai & Wesener, 2024 | PP754587 |
|
Thailand, Chantaburi Province, Tha Mai District, Wat Khao Sukim | |
| 34 | Zephronia macula Srisonchai & Wesener, 2024 | PX526095 | MZKKU (M400) | Thailand, Chantaburi Province, Tha Mai District, Wat Khao Sukim | |
| 35 | Zephronia macula Srisonchai & Wesener, 2024 | PX526097 | MZKKU (M437) | Thailand, Chachoengsao Province, Phanom Sarakham District, Wat Khao Hin Sorn | |
| 36 | Zephronia macula Srisonchai & Wesener, 2024 | PX526098 | MZKKU (M438) | Thailand, Rayong Province, Khao Chamao District, Wat Tham Suwan Phupha | |
| 37 | Zephronia macula Srisonchai & Wesener, 2024 | PX526100 | MZKKU (M457) | Thailand, Sa Kaeo Province, Khao Chakan District, Wat Tham Khao Chan | |
| 38 | Zephronia medongensis Zhao & Liu, 2022 | OP339793 | SCAU XZ01 | China, Xizang Autonomous Region, Medog County | |
| 39 | Zephronia ovalis Gray, 1832 | JX486068 | ZFMK Myr 0832 | Vietnam, Dong Nai Province, Cat Tien National Park | |
| 40 | Zephronia ovalis Gray, 1832 | PX526103 | MZKKU (M180) | Thailand, Si Sa Ket Province, Khunhan District, Huai Chan Waterfall | |
| 41 | Zephronia ovalis Gray, 1832 | PX526104 | MZKKU (M181) | Thailand, Si Sa Ket Province, Khunhan District, Huai Chan Waterfall | |
| 42 | Zephronia ovalis Gray, 1832 | PX526105 | MZKKU (M204) | Thailand, Si Sa Ket Province, Khunhan District, Huai Chan Waterfall | |
| 43 | Zephronia panhai Srisonchai, Sutcharit & Likhitrakarn, 2021 | OM509643 | ZFMK MYR 8118 | Thailand, Ratchaburi Province, Ratchaburi-Photharam Districts | |
| 44 | Zephronia panhai Srisonchai, Sutcharit & Likhitrakarn, 2021 | OM509644 | MHNG 3A | Thailand, Ratchaburi Province, Chom Bueang District, Tham Kao Bin Forest Park | |
| 45 | Zephronia panhai Srisonchai, Sutcharit & Likhitrakarn, 2021 | OM509645 | ZFMK MYR 8116 | Thailand, Ratchaburi Province, Ratchaburi-Photharam Districts | |
| 46 | Zephronia phrain Likhitrakarn & Golovatch, 2021 | OM509634 | ZFMK MYR 3499 | Thailand, Chiang Mai Province, Chiang Dao District, Padeng Lodge | |
| 47 | Zephronia phrain Likhitrakarn & Golovatch, 2021 | OM509635 | MYR3500 | Thailand, Chiangmai Province, Mueang District, Doi Suthep | |
| 48 | Zephronia phrain Likhitrakarn & Golovatch, 2021 | OM509636 | SMF | Thailand, Chiang Mai Province, Chiang Dao District, Tham Houay Luk | |
| 49 | Zephronia phrain Likhitrakarn & Golovatch, 2021 | OM509637 | SMF | Thailand, Chiang Mai Province, Chiang Dao District, Doi Chiang Dao, Ma Lee’s Resort | |
| 50 | Zephronia phrain Likhitrakarn & Golovatch, 2021 | OM509638 | SMF | Thailand, Chiang Mai Province, Chai Prakan District, Tham Ngam | |
| 51 | Zephronia phrain Likhitrakarn & Golovatch, 2021 | OM509639 | ZFMK MYR 4907 | Thailand, Chiang Mai Province, Chiang Dao | |
| 52 | Zephronia phrain Likhitrakarn & Golovatch, 2021 | OM509640 | MHNG 5G | Thailand, Lamphun Province, Mae Tha District, Doi Khuntan National Park | |
| 53 | Zephronia phrain Likhitrakarn & Golovatch, 2021 | OM509641 | MHNG 5I | Thailand, Chiang Mai Province, Chiang Dao District, Doi Chiang Dao | |
| 54 | Zephronia phrain Likhitrakarn & Golovatch, 2021 | OM509642 |
|
Thailand, Chiang Mai Province, Ban Musue | |
| 55 | Zephronia siamensis Hirst, 1907 (unverified) | JX486067.2 | FMNH-INS-72669 | Thailand, Chonburi Province, Sichang District, Koh Sichang | |
| 56 | Zephronia siamensis Hirst, 1907 | OR530089 | Thailand, Chonburi Province, Sichang District, Koh Sichang | ||
| 57 | Zephronia siamensis Hirst, 1907 | PP754592 |
|
Thailand, Chachoengsao Province, Phanom Sarakham District, Wat Khao Hin Sorn | |
| 58 | Zephronia siamensis Hirst, 1907 | PP754591 |
|
Thailand, Srakaeo Province, Khao Chakan District, Wat Tham Khao Chan | |
| 59 | Zephronia sp. (K45 Aow Noi Temple) | MW898741 |
|
Thailand, Prachuap Kiri Khan Province, Mueang District, Aow Noi Temple | |
| 60 | Zephronia sp. (Aow Noi Temple) | MW898742 | ZFMK MYR 8787 | Thailand, Prachuap Kiri Khan Province, Mueang District, Aow Noi Temple | |
| 61 | Zephronia sukhothaiensis Srimongkol & Srisonchai, 2026 | PX526107 | MZKKU (L43) | Thailand, Sukhothai Province, Ban Dan Lan Hoi District, Wat Khao Fang | |
| 62 | Zephronia sukhothaiensis Srimongkol & Srisonchai, 2026 | PX526108 | MZKKU (M323) | Thailand, Sukhothai Province, Ban Dan Lan Hoi District, Wat Khao Fang | |
| 63 | Zephronia sukhothaiensis Srimongkol & Srisonchai, 2026 | PX526110 | MZKKU (M325) | Thailand, Sukhothai Province, Ban Dan Lan Hoi District, Wat Khao Fang | |
| 64 | Zephronia tamarindina sp. nov. | PX737490 | MZKKU (L83) | Thailand, Phetchabun Province, Bueng Sam Phan District, Nimit Sat Cha Dhum Temple | This study |
| 65 | Zephronia tamarindina sp. nov. | PX737491 | MZKKU (L84) | Thailand, Phetchabun Province, Bueng Sam Phan District, Nimit Sat Cha Dhum Temple | This study |
| 66 | Zephronia tamarindina sp. nov. | PX737492 | MZKKU (UPNB01) | Thailand, Phetchabun Province, Bueng Sam Phan District, Nimit Sat Cha Dhum Temple | This study |
| 67 | Zephronia tamarindina sp. nov. | PX737493 | MZKKU (UPNB02) | Thailand, Phetchabun Province, Bueng Sam Phan District, Nimit Sat Cha Dhum Temple | This study |
| 68 | Zephronia tratensis Srimongkol & Srisonchai, 2026 | PX526112 | MZKKU (M415) | Thailand, Trat Province, Mueang District, Khao Khun Tham Monastery | |
| 69 | Zephronia tratensis Srimongkol & Srisonchai, 2026 | PX526113 | MZKKU (M416) | Thailand, Trat Province, Mueang District, Khao Khun Tham Monastery | |
| 70 | Zephronia triforma sp. nov. | PX737494 | MZKKU (L60) | Thailand, Nakhon Sawan Province, Takhli District, Buddhawajana Dhamma Retreat | This study |
| 71 | Zephronia triforma sp. nov. | PX737495 | MZKKU (L61) | Thailand, Nakhon Sawan Province, Takhli District, Buddhawajana Dhamma Retreat | This study |
| 72 | Zephronia triforma sp. nov. | PX737498 | MZKKU (UNSN01) | Thailand, Nakhon Sawan Province, Takhli District, Buddhawajana Dhamma Retreat | This study |
| 73 | Zephronia triforma sp. nov. | PX737499 | MZKKU (UNSN02) | Thailand, Nakhon Sawan Province, Takhli District, Buddhawajana Dhamma Retreat | This study |
| 74 | Zephronia triforma sp. nov. | PX737496 | MZKKU (L227) | Thailand, Phitsanulok Province, Wat Bot District, Ban Nam Kob | This study |
| 75 | Zephronia triforma sp. nov. | PX737497 | MZKKU (L229) | Thailand, Phitsanulok Province, Wat Bot District, Ban Nam Kob | This study |
| 76 | Zephronia triforma sp. nov. | PX737500 | MZKKU (UPLK07) | Thailand, Phitsanulok Province, Wat Bot District, Khao Bo Ma Viewpoint | This study |
| 77 | Zephronia triforma sp. nov. | PX737501 | MZKKU (UPLK12) | Thailand, Phitsanulok Province, Wat Bot District, Khao Bo Ma Viewpoint | This study |
| 78 | Zephronia viridisoma Rosenmejer & Wesener, 2021 | MW898739 |
|
Thailand, Nakhon Si Thammarat Province, Sichon District, Khao Lark Waterfall | |
| 79 | Zephronia zhouae Zhao & Liu, 2022 | OP339794 | SCAU YN02 | China, Yunnan Province, Diqing Tibetan Autonomous Prefecture, Weixi County, Laowo Village | |
| Outgroups | |||||
| 80 | S. aesculus Rosenmejer & Wesener, 2021 | MW898737 |
|
Thailand, Phuket Province, Kathu District, Forest | |
| 81 | S. cf. aesculus Rosenmejer & Wesener, 2021 | MW898738 |
|
Thailand, Nakhon Si Thammarat Province, Khao Luang NP | |
| 82 | S. benquii Liu & Wesener, 2022 | OP339792 | SCAU MMY01 | China, Guizhou, Tongren City, Jiangkou County, Guanhe Town, Guanhe Village, Maomaoyan | |
| 83 | S. bolavensis Wesener, 2019 | MK330982 | MHNG LT-10/24 | Laos, Champasak Province, Bolaven Plateau, 3 km S of Ban Nong Luang, Tad Kameud | |
| 84 | S. denticulatum Wesener, 2019 | MK330984 | MHNG LT-10/12 | Laos, Oudomxai Province, ca 3 km E of Tad Lak 11, SE of Oudomxai city | |
| 85 | S. huzhengkuni Zhao, Yu & Liu, 2020 | MT657327 | SCAU SP02 | China, Guizhou Province, Tongren City, Fanjingshan National Nature Reserve | |
| 86 | S. huzhengkuni Zhao, Yu & Liu, 2020 | MT657328 | SCAU SP03 | China, Guizhou Province, Tongren City, Fanjingshan National Nature Reserve | |
| 87 | S. lachneeis Wesener, 2019 | MK330983 | MHNG LT-10/12 | Laos, Oudomxai Province, ca 3 km E of Tad Lak 11, SE of Oudomxai city | |
| 88 | S. laoticum Wesener, 2019 | MK330975 | SMF | Laos, Vientiane Province, Vang Vieng | |
| 89 | S. meridionalis Bhansali & Wesener, 2022 | OM509648 | MHNG 4B-2 | Thailand, Yala Province, Bannang Sata District, Bang Lang National Park, near Than To Waterfall | |
| 90 | S. nigrum Wesener, 2019 | MK330976 | SMF | Laos, Champasak Province, Muang Bachieng, Ban Lak 35, Tad Etu | |
| 91 | S. nonghinensis Srikampha & Srisonchai, 2025 | PQ838982 | MZKKU (A35) | Thailand, Loei Province, Nong Hin District, Suan Hin Pha Ngam (Hin Pha Ngam Park) | |
| 92 | S. nonghinensis Srikampha & Srisonchai, 2025 | PQ838983 | MZKKU (A36) | Thailand, Loei Province, Nong Hin District, Suan Hin Pha Ngam (Hin Pha Ngam Park) | |
| 93 | S. nonghinensis Srikampha & Srisonchai, 2025 | PQ838984 | MZKKU (E28) | Thailand, Loei Province, Nong Hin District, Suan Hin Pha Ngam (Hin Pha Ngam Park) | |
| 94 | S. nonghinensis Srikampha & Srisonchai, 2025 | PQ838986 | MZKKU (M237) | Thailand, Loei Province, Nong Hin District, Suan Hin Pha Ngam (Hin Pha Ngam Park) | |
| 95 | S. onyx Srikampha & Srisonchai, 2025 | PQ838987 | MZKKU (M211) | Thailand, Buang Kan Province, Seka District, Wat Tham Chum Chang (Nam Yaad Clift) | |
| 96 | S. peterjaegeri Wesener, 2019 | MK330972 | SMF SD553 | Laos, Luang Prabang Province, SE Luang Prabang, Nam Khan, Ban Pak Bak, Houay Kho | |
| 97 | S. petramurum Srikampha & Srisonchai, 2025 | PQ838989 | MZKKU (E5) | Thailand, Khon Kaen Province, Phu Pha Man District, Ban Khao Wong | |
| 98 | S. petramurum Srikampha & Srisonchai, 2025 | PQ838991 | MZKKU (L128) | Thailand, Khon Kaen Province, Phu Pha Man District, Ban Khao Wong | |
| 99 | S. petramurum Srikampha & Srisonchai, 2025 | PQ838992 | MZKKU (L6) | Thailand, Chaiyaphum Province, Khon San District, Mortagek Monastery | |
| 100 | S. petramurum Srikampha & Srisonchai, 2025 | PQ838993 | MZKKU (M217) | Thailand, Khon Kaen Province, Phu Pha Man District, Ban Khao Wong | |
| 101 | S. phouloei Wesener, 2019 | MK330974 | ZMUC00040257 | Laos, Houaphan Province, Phou Loei | |
| 102 | S. schwendingeri Wesener, 2019 | MK330978 | MHNG LT-10/03 | Laos, Vientiane Province, trail to Tham Pou Kham, W. of Vang Vieng | |
| 103 | S. schwendingeri Wesener, 2019 | MK330981 | SMF | Laos, Vientiane Province, Vang Vieng, Tham Pou Kham | |
| 104 | Sphaerobelum sp. L07 | MK330979 | ZMUC00040261 | Laos, Khammouane Province, Ban Khounkham [Khun Kham] (Nahin) | |
| 105 | Sphaerobelum sp. L10 | MK330980 | SMF | Laos, Vientiane Province, Vang Vieng, W. of Nam Song, Tham Nam Or Khem | |
| 106 | S. spinatum Wesener, 2019 | MK330973 | ZMUC00040258 | Laos, Vientiane Province, Phou Khao Khouay | |
| 107 | S. truncatum Wongthamwanich, 2012 | JN885184 | FMNH-INS 0000 072 674 | Thailand, Nan Province, Song Khwae District, Na Rai Luang Subdistrict, Pang Hi Village | |
| 108 | S. tujiaphilum Zhao & Liu, 2022 | OP339785 | SCAU BHS01 | China, Guizhou, Tongren City, Jiangkou County, Taiping Town, Baiheshan Village | |
| 109 | S. tujiaphilum Zhao & Liu, 2022 | OP339787 | SCAU JXT02 | China, Guizhou, Tongren City, Jiangkou County, Taiping Town, Jiang-xitun Village | |
| 110 | S. tujiaphilum Zhao & Liu, 2022 | OP339788 | SCAU SD03 | China, Guizhou, Tongren City, Jiangkou County, Guanhe Town, Sidu Village | |
| 111 | S. tujiaphilum Zhao & Liu, 2022 | OP339789 | SCAU DW01 | China, Guizhou, Tongren City, Jiangkou County, Dewang Town, Xiaobang Village | |
| 112 | S. turcosa Srisonchai & Pimvichai, 2023 | OR530087 |
|
Thailand, Loei Province, Mueang Loei District, Phu Pha Lom Forest Park | |
| 113 | S. turcosa Srisonchai & Pimvichai, 2023 | OR530087 |
|
Thailand, Loei Province, Mueang Loei District, Phu Pha Lom Forest Park | |
| 114 | Arthrosphaera brandtii (Humbert, 1865) | FJ409915 | FMNH-INS 8650 | Tanzania, Usambara hills | |
| 115 | Cryxus ovalis (Linnaeus, 1758) (unverified) | JX486069.2 | ZFMK MYR0824 | Vietnam, Dong Nai Province, Cat Tien National Park | |
| 116 | Epicyliosoma sp. | AF218270 | NA | NA | |
| 117 | Glomeris marginata (Villers, 1789) | FJ409909 | ZFMK Myr009 | Germany, Bonn, Venusberg | |
| 118 | P. inthanonense Donworth & Wesener, 2024 | PP297645 | MHNG 4E-2 | Thailand, Chiang Mai Province, Chom Thong District, Doi Inthanon National Park | |
| 119 | P. inthanonense Donworth & Wesener, 2024 | PP297646 | MHNG 7A | Thailand, Chiang Mai Province, Chom Thong District, Doi Inthanon National Park | |
| 120 | P. naevium Donworth & Wesener, 2024 | PP297647 | MHNG 4B-1 | Thailand, Yala Province, Than To District, Bang Lang National Park, Than To Waterfall | |
| 121 | P. naevium Donworth & Wesener, 2024 | PP297648 |
|
Thailand, Yala Province, Than To District, Bang Lang National Park | |
| 122 | P. naevium Donworth & Wesener, 2024 | PP297649 |
|
Thailand, Yala Province, Than To District, Bang Lang National Park | |
| 123 | P. naevium Donworth & Wesener, 2024 | PP297650 |
|
Thailand, Yala Province, Than To District, Bang Lang National Park | |
| 124 | Sphaeromimus splendidus Wesener & Sierwald, 2005 | FJ409917 | FMMC-INS 6702 | Madagascar, Sainte Luce S9 | |
The chromatograms of all sequences were examined, aligned, and trimmed using BioEdit v. 7.0.5.3 (https://bioedit.software.informer.com/download/) (
COI barcoding trees were reconstructed using maximum likelihood (ML) and Bayesian inference (BI) methods. The ML tree was generated using the IQ-tree on XSEDE 1.6.6 (
Interspecific and intraspecific genetic distances were calculated using the uncorrected p-distance model in MEGA X (
The 658 bp COI gene fragments from 124 sequences contained 376 polymorphic sites, 282 conservative sites, and 316 parsimony informative sites. The genetic distances (percentage) between Zephronia and other closely related genera were from 6.85–25.27% between Sphaerobelum and Zephronia, 16.44–23.29% between Prionobelum and Zephronia, and 18.02–23.38% between Cryxus Leach, 1814 and Zephronia.
Within Zephronia itself, interspecific divergence ranged from 8.68 to 24.20%, with the lowest distance observed between Z. siamensis and Z. dawydoffi. Pairwise interspecific distances between each taxon and its closest congeners within Zephronia were 8.68–21.92% in Z. dawydoffi, 12.79–22.98% in Z. laotica, 10.81–22.07% in Z. tamarindina sp. nov., and 10.81–22.83% in Z. triforma sp. nov. The intraspecific genetic distances reached up to 5.18% in Z. dawydoffi, 5.95% in Z. laotica, 0.61% in Z. tamarindina sp. nov., and 5.63% in Z. triforma sp. nov., respectively. Notably, Z. dawydoffi in this study differed by 3.65–5.18% from the GenBank sequence (MK330971), while Z. laotica differed by 4.88–5.95% from the GenBank sequence (MK330977) (see Suppl. material
COI barcoding tree reconstruction using BI and ML methods produced partially incongruent topologies. However, most interspecific relationships, particularly among Zephronia s.s. from Thailand, Laos, Vietnam, and Cambodia, were broadly consistent (Fig.
COI barcoding trees based on Bayesian Inference (BI) and maximum likelihood (ML) of the genus Zephronia and related genera. Numbers on each node represent posterior probabilities (PP) from BI and bootstrap support (BS) from ML. Background colours indicate the two new species (Zephronia tamarindina sp. nov. and Zephronia triforma sp. nov.), and the two new recorded species from Thailand (Zephronia dawydoffi Attems, 1953 and Zephronia laotica Wesener, 2019), relating to the photograph of live specimens. The scale bar represents the branch length. Colour of scientific letters: blue = outgroup species, black = Zephronia species.
Overall, nodal support across deeper branches was generally low, limiting the resolution of relationships among Zephronia, Sphaerobelum, Prionobelum, and Cryxus. Both BI and ML trees indicated that Zephronia is non-monophyletic. In the BI tree, Zephronia, Sphaerobelum, Prionobelum, and Cryxus were intermixed, forming a poorly resolved polytomy with low support (PP = 0.59). Similarly, in the ML, Zephronia was divided into four major clades with weak support (BS = 47). While most of the Zephronia s.s. clustered together, Z. phrain, Z. viridisoma, and Zephronia sp. (Aow Noi Temple) formed a separate clade. In addition, Z. hui, Z. medogensis, and Z. zhouae were interspersed with Sphaerobelum in two distinct clades.
All other Zephronia s.s. specimens from Thailand, Laos, Vietnam, and Cambodia, including the newly sequenced Z. dawydoffi, Z. laotica, Z. tamarindina sp. nov., and Z. triforma sp. nov., formed a moderately to well-supported monophyletic group in both BI and ML trees (BS = 74, PP = 0.91). Within this clade, the two new species were nested among members of the Zephronia s.s. group (Z. chrysomallos, Z. erawani, Z. golovatchi, Z. lannaensis, and Z. panhai) with strong support (BS = 99, PP = 1.00). Z. tamarindina sp. nov. and Z. triforma sp. nov. were recovered as sister species (BS = 99, PP = 1.00). Z. dawydoffi and Z. laotica were grouped together with Z. chantaburiensis, Z. macula, Z. ovalis, and Z. siamensis with strong support (BS = 79, PP = 1.00). Sequences of Z. dawydoffi and Z. laotica from this study clustered with their respective GenBank references (MK330971 of Z. dawydoffi: BS = 50, PP = 1.00; MK330977 of Z. laotica: BS = 100, PP = 1.00).
Class Diplopoda de Blainville in Gervais, 1844
Order Sphaerotheriida Brandt, 1833
Family Zephroniidae Gray, 1843
Subfamily Zephroniinae Gray, 1843
Tribe Zephroniini Gray, 1843
Zephronia ovalis Gray, 1832.
59 species, including the two new species described here.
Cambodia, China (including Hong Kong), India (NE), Indonesia, Laos, Malaysia, Myanmar, Nepal, Singapore, Thailand, and Vietnam.
Zephronia is one of the most species-rich genera within Zephroniidae, currently comprising 59 described species. Many species remain insufficiently studied, especially type re-examinations are lacking, with many known only from females, particularly those from Myanmar, Borneo, Sumatra, and India. Consequently, the genus is in urgent need of a revision, and some species currently placed in Zephronia may be transferred to separate genera in the future.
Zephronia differs from other genera of Zephroniinae by the following combination of characters: endotergum weakly modified; posterior margin usually flat (rarely modified); outer area without setae; marginal bristles arranged in 1–5 rows; middle area often with a single row of circular cuticular impressions; tarsi of leg pairs 5–21 usually with more than one apical spine (rarely with a single apical spine) and several ventral spines; male anterior telopods with four podomeres distal to syncoxite; telopoditomere 2 bearing a large, curved process forming a clamp-like structure; telopoditomeres 3 and 4 simple; male posterior telopods with four telopoditomeres; process of telopoditomere 2 slender and apically curved; telopoditomeres 3 and 4 with a membranous ledge; telopoditomere 3 with a row of large, crenulated, sclerotised teeth; telopoditomere 4 with one or two sclerotised spines; female vulvae conspicuous and slender, operculum usually rounded.
See more recent diagnoses for Zephronia sensu lato and Zephronia sensu stricto in
Zephronia s.s. currently includes Z. chantaburiensis, Z. chrysomallos, Z. dawydoffi, Z. enghoffi, Z. erawani, Z. golovatchi, Z. hui Liu & Wesener, 2022, Z. konkakinhensis Semenyuk, Golovatch & Wesener, 2018, Z. lannaensis, Z. laotica, Z. macula, Z. medogensis Zhao & Liu, 2022, Z. montis Semenyuk, Golovatch & Wesener, 2018, Z. ovalis, Z. panhai, Z. phrain, Z. siamensis, Z. sukhothaiensis, Z. tamarindina sp. nov., Z. tratensis, Z. triforma sp. nov., Z. zhouae Zhao & Liu, 2022.
Among the examined material, colour variation initially suggested the presence of multiple distinct taxa. In particular, one species, Z. triforma sp. nov., exhibits three distinct colour morphs, which could be misinterpreted as representing separate species based on external appearance alone. In contrast, Z. tamarindina sp. nov. and the two previously described species each show a single, consistent colour form. However, integrative analyses combining morphological characters and COI sequence data confirm that the three colour morphs belong to a single species, Z. triforma sp. nov., whereas the remaining populations correspond to two new species and two previously described species.
In this study, we focus on describing the two new species and the two newly recorded species from Thailand.
Zephronia dawydoffi Attems, 1953: 156.
Zephronia dawydoffi —
• 18 ♂♂ 33 ♀♀ (MZKKU); Thailand, Khon Kaen Province, Mueang District, Thung Setthi Temple; 16°24'33"N, 102°53'05.7"E; ca. 150 m a.s.l.; 12 July 2022; leg. C. Wilaisorn and MZKKU students; • 5 ♂♂ 7 ♀♀ (MZKKU); Thailand, Buri Ram Province, Khu Mueang District, Dong Kheng rubber plantation; 15°19'46"N, 103°00'59"E; ca. 159 m a.s.l.; 6 July 2022; leg. C. Wilaisorn and MZKKU students; • 4 ♂♂ 2 ♀♀ (MZKKU); Thailand, Buri Ram Province, Mueang District, Khao Kradong; 14°56'21.5"N, 103°05'28.8"E; ca. 213 m a.s.l.; 10 September 2023; leg. C. Wilaisorn and MZKKU students; • 3 ♀♀ (MZKKU); Thailand, Buri Ram Province, Mueang District, Ban Khok Ta Sing Waterfall; 14°55'22.8"N, 103°06'57.9"E; ca. 180 m a.s.l.; 5 June 2022; leg. C. Wilaisorn and MZKKU students. • 12 ♂♂ 21 ♀♀ (MZKKU); Thailand, Buri Ram Province, Khu Mueang District, Non Somboon Monastery; 15°16'32"N, 102°59'13"E; ca. 200 m a.s.l.; 31 July 2022; leg. C. Wilaisorn and MZKKU students.
Zephronia dawydoffi Attems, 1953, ♂ from Thung Setthi Temple, Khon Kaen Province, Thailand (MZKKU) — SEM. A. Collum, dorsal view; B. Head, anterior view; C. Antenna, lateral view; D. Tömösváry’s organ; E, F. Apical cones, lateral view; G, J. Gnathochilarium, anterior and ventral views, respectively; H. Sensory cones on inner palpi, ventral view; I. Sensory cones on central pad, ventral view; K. Epipharynx, ventral view; L. Right mandible, mesal view. Abbreviations: 3it = 3-combed inner tooth, co = condylus, Cp = central pad, ct = central tooth, et = external tooth of epipharynx, Et = external tooth of mandible, Ia = inner area, il = incisura lateralis, ip = inner palpi, ll = lamellae linguales, me = mentum, mp = molar plate, o = ommatidia, pl = pectinate lamellae, rsp = row of spines, sc = sensory cone, st = stipites, t = Tömösváry’s organ. Scale bars in millimetres.
Zephronia dawydoffi Attems, 1953, ♂ and ♀ from Thung Setthi Temple, Khon Kaen Province, Thailand (MZKKU) — SEM. A–C. Surface of midbody tergite, all in dorsal views (overview, anterior margin, surface, respectively); D. Subanal plate; E. Anal shield, underside, ventral view; F. Locking carinae; G. Surface of anal shield; H. First pleurite; I. Midbody pleurite. Scale bars in millimetres.
Zephronia dawydoffi Attems, 1953, ♂ and ♀ from Thung Setthi Temple, Khon Kaen Province, Thailand (MZKKU) — SEM. A. Legs 1; B. Legs 2 with gonopores; C. Leg 2 with vulvae; D. Midbody legs (arrow points to femoral ridge); E. Small triangular spines on femur; F. Coxal process on midbody legs. Abbreviation: bu = bursa, cx = coxa, fe = femur, op = operculum, po = postfemur, pre = prefemur, rdg = femoral ridge, st-pl = stigmatic plate, ta = tarsus, ti = tibia. Scale bars in millimetres.
Zephronia dawydoffi Attems, 1953, ♂from Thung Setthi Temple, Khon Kaen Province, Thailand (MZKKU) — SEM. A–C. Overview of telopods (anterior, posterior, and ventral views, respectively); D. Crenulated teeth of posterior telopods, posterior view; E. Sclerotised spots on immovable finger of posterior telopod, sublateral view; F. Anterior telopod, ventral view; G. Inner horns, anterior view. Abbreviations: cr-t = crenulated teeth, ml = membranous lobe, scl-s = sclerotised spots, syn-cx = syncoxite. Scale bars in millimetres.
In Z. dawydoffi, the Tömösváry’s organ is positioned at the brim rather than inside the antennal groove (Fig.
CAMBODIA (Ream, Sre Umbell; Kampot, Kampong Speu).
Measurements. Males: body length = 10.5–18.8 mm; width (broadest), of thoracic shield = 5.3–8.6 mm, of tergite 7 = 5.6–9.3 mm; highest of thoracic shield = 3.1–5.5 mm, of tergite 7 = 3.3–6.9 mm. Females: body length = 11.5–22.0 mm; width (broadest), of thoracic shield = 5.8–10.6 mm, of tergite 7 = 6.4–11.7 mm; highest of thoracic shield = 4.0–7.1 mm, of tergite 7 = 4.4–8.2 mm.
Colouration (Fig.
Head (Fig.
Antennae (Fig.
Epipharynx (Fig.
Gnathochilarium (Fig.
Mandibles (gnathal lobe) (Fig.
Tegument (Figs
Collum (Fig.
Thoracic shield. Anterior margin with shallow canal covered with a few scattered long setae, with 5–7 small keels.
Midbody tergite (Fig.
Anal shield (Fig.
Endotergum of thoracic shield and midbody tergite (Fig.
Pleurite (laterotergite) (Fig.
Subanal plate of female (Fig.
Stigmatic plates (Fig.
Legs (Fig.
Male sexual characters (Fig.
Anterior telopods (Figs
Posterior telopods (Figs
Female sexual characters (Fig.
Variation in body colouration and pattern was observed among specimens. Most individuals displayed dark green tergites, while others exhibited greenish-brown or dark brown tones. Typically, two yellow patches were present on dorsal tergites, though in some specimens these patches appeared faded or paler in colour. The patch shapes also varied, most commonly forming two semicircular markings (Fig.
Morphological differences were observed between Thai specimens and the original description by
Zephronia dawydoffi in Thailand is currently known from two provinces in the northeastern region: four localities in the Buri Ram Province and only one locality in the Khon Kaen Province (Fig.
Compared with its type locality in southern Cambodia, the known distribution of Z. dawydoffi is extended into Thailand, representing a geographic range spanning approximately 300–700 km.
Zephronia laotica Wesener, 2019: 205.
Zephronia laotica —
• 10 ♂♂ 7 ♀♀ (MZKKU); Thailand, Si Sa Ket Province, Khun Han District, Mae Lamoon Durian plantation; 14°33'00"N, 104°31'51"E; ca. 228 m a.s.l.; 29 July 2022; leg. C. Wilaisorn and MZKKU students; • 10 ♂♂ 12 ♀♀ (MZKKU); Thailand, Si Sa Ket Province, Khun Han District, Kantrom Tai Temple; 14°34'32"N, 104°20'33"E; ca. 188 m a.s.l.; 6 October 2024; leg. C. Wilaisorn and MZKKU students; • 2 ♀♀ (MZKKU); Thailand, Si Sa Ket Province, Khun Han District, Ta Nuan Rubber plantation; 14°33'47.9"N, 104°21'24.2"E; ca. 180 m a.s.l.; 10 June 2023; leg. C. Wilaisorn and MZKKU students.
The position of the Tömösváry’s organ is located at the brim rather than inside the antennal groove (Fig.
Zephronia laotica Wesener, 2019, ♂ from Mae Lamoon Durian plantation, Si Sa Ket Province, Thailand (MZKKU) — SEM. A. Collum, dorsal view; B. Head, anterior view; C. Antenna, lateral view; D. Tömösváry’s organ; E, F. Apical cones, lateral view; G, J. Gnathochilarium, anterior and ventral views, respectively; H. Sensory cones on inner palpi, ventral view; I. Sensory cones on central pad, ventral view; K. Epipharynx, ventral view; L. Right mandible, mesal view. Abbreviations: 3it = 3-combed inner tooth, co = condylus, Cp = central pad, ct = central tooth, et = external tooth of epipharynx, Et = external tooth of mandible, Ia = inner area, il = incisura lateralis, ip = inner palpi, ll = lamellae linguales, me = mentum, mp = molar plate, o = ommatidia, pl = pectinate lamellae, rsp = row of spines, sc = sensory cone, st = stipites, t = Tömösváry’s organ. Scale bars in millimetres.
Zephronia laotica Wesener, 2019, ♂ and ♀ from Mae Lamoon Durian plantation, Si Sa Ket Province, Thailand (MZKKU) — SEM. A–C. Surface of midbody tergite, all in dorsal views (overview, anterior margin, surface, respectively); D. Subanal plate; E. Anal shield, underside, ventral view; F. Locking carinae (arrowheads point to carinae); G. Surface of anal shield; H. First pleurite; I. Midbody pleurite. Scale bars in millimetres.
Zephronia laotica Wesener, 2019, ♂ and ♀ from Mae Lamoon Durian plantation, Si Sa Ket Province, Thailand (MZKKU) — SEM. A. Legs 1; B. Legs 2 with gonopores; C. Leg 2 with vulvae; D. Midbody legs (arrow points to femoral ridge); E. Small triangular spines on femur; F. Coxal process on midbody legs. Abbreviation: bu = bursa, cx = coxa, fe = femur, op = operculum, po = postfemur, pre = prefemur, rdg = femoral ridge, st-pl = stigmatic plate, ta = tarsus, ti = tibia. Scale bars in millimetres.
Zephronia laotica Wesener, 2019, ♂ from Mae Lamoon Durian plantation, Si Sa Ket Province, Thailand (MZKKU) — SEM. A–C. Overview of telopods (anterior, posterior, and ventral views, respectively); D. Crenulated teeth of posterior telopods, posterior view; E. Sclerotised spots on immovable finger of posterior telopod, sublateral view; F. Anterior telopod, ventral view; G. Inner horns, anterior view. Abbreviations: cr-t = crenulated teeth, ml = membranous lobe, scl-s = sclerotised spots. Scale bars in millimetres.
LAOS, Champasak Province, east of Mekong, Garden of Erawan Riverside Hotel.
Measurements. Males: body length = 10.2–15.2 mm; width (broadest), of thoracic shield = 5.2–7.9 mm, of tergite 7 = 5.5–8.1 mm; highest of thoracic shield = 4.4–6.6 mm, of tergite 7 = 3.8–6.1 mm. Females: body length = 11.4–15.5 mm; width (broadest), of thoracic shield = 5.8–10.6 mm, of tergite 7 = 6.2–11.1 mm; highest of thoracic shield = 4.0–7.8 mm, of tergite 7 = 4.3–8.1 mm.
Colouration (Fig.
Head (Fig.
Antennae (Fig.
Epipharynx (Fig.
Gnathochilarium (Fig.
Mandibles (gnathal lobe) (Fig.
Tegument (Figs
Collum (Fig.
Thoracic shield. Anterior margin with shallow canal covered by numerous long setae; with 5–8 conspicuous keels.
Midbody tergite (Fig.
Anal shield (Fig.
Endotergum of thoracic shield and midbody tergite (Fig.
Pleurite (laterotergite) (Fig.
Subanal plate of female (Fig.
Stigmatic plates (Fig.
Legs (Fig.
Male sexual characters (Fig.
Anterior telopods (Figs
Posterior telopods (Figs
Female sexual characters (Fig.
Noticeable variations occur in colouration and patterns of tergites. Most specimens display two yellow patches on dorsal tergites, typically semicircular or rectangular in shape (Fig.
The Thai specimens differ from the original description by
In contrast to the type specimens, the paratergites are more noticeably protruding posteroventrad, and the thoracic shield comprises 5–8 keels. The legs of the Thai specimens differ slightly from what was originally described. Thai specimens possessed 12–16 small triangular spines on their femurs, while no spines were mentioned in the original description. Because these spines are minute and the original description was based on light microscopy only, their absence from
Despite this obvious variation, the male anterior and posterior telopods, the principal species-diagnostic structures in Zephronia, are indistinguishable between the Thai and Lao material. The Thai sequences cluster with the topotypic sequence MK330977 with maximal support (BS = 100, PP = 1.00) at a divergence of 4.88–5.95%, below the lowest interspecific distance recovered in this study (8.68%). We therefore treat the Thai population as a geographically divergent population of Z. laotica, while acknowledging that denser sampling in Laos and nuclear markers may reveal cryptic species-level structure.
Zephronia laotica is currently recorded only from the lower northeastern region of Thailand, specifically Si Sa Ket Province (Fig.
Holotype • ♂ (MZKKU-MYR0021); Thailand, Phetchabun Province, Bueng Sam Phan District, Nimit Sat Cha Tham Temple; 15°49'16.8"N, 100°55'03.9"E; ca 204 m a.s.l.; 3 September 2023; leg. C. Wilaisorn and MZKKU students. Paratypes. • 4 ♂♂ 4 ♀♀ (MZKKU-MYR0022); same data as for holotype; • 1 ♂ 1 ♀ (
The position of the Tömösváry’s organ located at the brim rather than inside the antennal groove (Fig.
Zephronia tamarindina sp. nov., ♂ paratype (MZKKU) — SEM. A. Collum, dorsal view; B. Head, anterior view; C. Antenna, lateral view; D. Tömösváry’s organ; E, F. Apical cones, lateral view; G, J. Gnathochilarium, anterior and ventral views, respectively; H. Sensory cones on inner palpi, ventral view; I. Sensory cones on central pad, ventral view; K. Epipharynx, ventral view; L. Right mandible, mesal view. Abbreviations: 3it = 3-combed inner tooth, co = condylus, Cp = central pad, ct = central tooth, et = external tooth of epipharynx, Et = external tooth of mandible, Ia = inner area, il = incisura lateralis, ip = inner palpi, ll = lamellae linguales, me = mentum, mp = molar plate, o = ommatidia, pl = pectinate lamellae, rsp = row of spines, sc = sensory cone, st = stipites, t = Tömösváry’s organ. Scale bars in millimetres.
Zephronia tamarindina sp. nov., ♂ paratype (MZKKU), ♀ paratype (MZKKU) — SEM. A–C. Surface of midbody tergite, all in dorsal views (overview, anterior margin, surface, respectively); D. Subanal plate; E. Anal shield, underside, ventral view; F. Locking carinae; G. Surface of anal shield; H. First pleurite; I. Midbody pleurite. Scale bars in millimetres.
Zephronia tamarindina sp. nov., ♂ paratype (MZKKU), ♀ paratype (MZKKU) — SEM. A. Legs 1; B. Legs 2 with gonopores; C. Leg 2 with vulvae; D. Midbody legs (arrow points to femoral ridge); E. Small triangular spines on femur; F. Coxal process on midbody legs. Abbreviation: bu = bursa, cx = coxa, fe = femur, Mp = membranous process, op = operculum, po = postfemur, pre = prefemur, rdg = femoral ridge, st-pl = stigmatic plate, ta = tarsus, ti = tibia. Scale bars in millimetres.
Zephronia tamarindina sp. nov., ♂ paratype (MZKKU) — SEM. A–C. Overview of telopods (anterior, posterior, and ventral views, respectively); D. Crenulated teeth of posterior telopods, posterior view; E. Anterior telopod, ventral view; F. Crenulated teeth of anterior telopods; G. Sclerotised spots on immovable finger of posterior telopod, sublateral view; H. Inner horns, anterior view. Abbreviations: cr-t = crenulated teeth, ml = membranous lobe, scl-s = sclerotised spots. Scale bars in millimetres.
Measurements. Male holotype; body length = 18.6 mm; width (broadest), of thoracic shield = 9.5 mm, of tergite 7 = 10.5 mm; highest of thoracic shield = 6.1 mm, of tergite 7 = 7.8 mm. Males: body length = 15.0–22.5 mm; width (broadest), of thoracic shield = 7.3–11.7 mm, of tergite 7 = 7.5–12.3 mm; highest of thoracic shield = 5.8–7.6 mm, of tergite 7 = 6.9–8.8 mm. Females: body length = 14.8–26.2 mm; width (broadest), of thoracic shield = 7.1–12.6 mm, of tergite 7 = 6.6–13.7 mm; highest of thoracic shield = 4.8–8.7 mm, of tergite 7 = 8.8–10.0 mm.
Colouration (Fig.
Head (Fig.
Antennae (Fig.
Epipharynx (Fig.
Gnathochilarium (Fig.
Mandibles (gnathal lobe) (Fig.
Tegument (Figs
Collum (Fig.
Thoracic shield. Anterior margin with a shallow canal covered by numerous long setae; with 5–9 conspicuous keels.
Midbody tergite (Fig.
Anal shield (Fig.
Endotergum of thoracic shield and midbody tergite (Fig.
Pleurite (laterotergite) (Fig.
Subanal plate of female (Fig.
Stigmatic plates (Fig.
Legs (Fig.
Male sexual characters (Fig.
Anterior telopods (Figs
Posterior telopods (Figs
Female sexual characters (Fig.
The variation in colouration of paratergites was observed, with most specimens exhibiting an olive green colour, while some appeared orangish brown. Juveniles and smaller individuals typically showed olive greenish-brown colouration with rectangular patches on tergites. A distinctive feature of this species is the conspicuous triangular membranous process present on the coxae of male leg-pair 2 (Fig.
Zephronia tamarindina sp. nov. is currently known only from its type locality in Phetchabun Province and is thus considered endemic to this area (Fig.
The name is an adjective that derives from Tamarindus (tamarind), an important fruit cultivated in Phetchabun Province, the type locality. The name is treated as a feminine adjective in reference to the species body colouration, which changes from pale olive green in juveniles to a darker orangish brown in adults, resembling the ripening process of tamarind fruit from green to brown.
Holotype • ♂ (MZKKU-MYR0023); Thailand, Phitsanulok Province, Wat Bot District, Ban Nam Kob; 16°59'43.6"N, 100°23'34.0"E; ca 95 m a.s.l.; 6 August 2024; leg. C. Wilaisorn and MZKKU students. Paratypes. • 29 ♂♂ 25 ♀♀ (MZKKU-MYR0024), same data as for holotype; • 1 ♂ 1 ♀ (
• 13 juveniles (MZKKU), same data as for holotype; • 29 ♂♂, 13 ♀♀ (MZKKU); Thailand, Phitsanulok Province, Wat Bot District, Khao Bo Ma Viewpoint; 17°18'20.3"N, 100°26'29.2"E; ca. 216 m a.s.l.; 10 July 2024; leg. C. Wilaisorn, R. Srisonchai and MZKKU students; • 7 ♂♂ 4 ♀♀ (MZKKU); Thailand, Nakhon Sawan Province, Takhli District, Buddhawajana Dhamma Retreat; 15°17'51.7"N, 100°22'24.6"E; ca 70 m a.s.l.; 31 July 2023; leg. C. Wilaisorn, R. Srisonchai, and MZKKU students.
The position of the Tömösváry’s organ is located at the brim rather than inside the antennal groove (Fig.
Photograph of live specimens of Zephronia triforma sp. nov. and habitats. A, D, G. Male paratypes (MZKKU); B, E, H. Female paratypes (MZKKU); C, F, I. Live specimens of a population from Nakhon Sawan Province (MZKKU); J. Dipterocarp forest at the type locality; K. Dry deciduous forest at Nakhon Sawan sampling area. Scale bars: 5.0 mm.
Zephronia triforma sp. nov., ♂ paratype (MZKKU) — SEM. A. Collum, dorsal view; B. Head, anterior view; C. Antenna, lateral view; D. Tömösváry’s organ; E, F. Apical cones, lateral view; G, J. Gnathochilarium, anterior and ventral views, respectively; H. Sensory cones on inner palpi, ventral view; I. Sensory cones on central pad, ventral view; K. Epipharynx, ventral view; L. Right mandible, mesal view. Abbreviations: 3it = 3-combed inner tooth, co = condylus, Cp = central pad, ct = central tooth, et = external tooth of epipharynx, Et = external tooth of mandible, Ia = inner area, il = incisura lateralis, ip = inner palpi, ll = lamellae linguales, me = mentum, mp = molar plate, o = ommatidia, pl = pectinate lamellae, rsp = row of spines, sc = sensory cone, st = stipites, t = Tömösváry’s organ. Scale bars in millimetres.
Zephronia triforma sp. nov., ♂ paratype (MZKKU), ♀ paratype (MZKKU) — SEM. A–C. Surface of midbody tergite, all in dorsal views (overview, anterior margin, surface, respectively); D. Subanal plate; E. Anal shield, underside, ventral view; F. Locking carinae; G. Surface of anal shield; H. First pleurite; I. Midbody pleurite. Scale bars in millimetres.
Zephronia triforma sp. nov., ♂ paratype (MZKKU), ♀ paratype (MZKKU) — SEM. A. Legs 1; B. Legs 2 with gonopores; C. Leg 2 with vulvae; D. Midbody legs (arrow points to femoral ridge); E. Phoretic deutonymphs of an unknown mite; F. Small triangular spines on femur. Abbreviation: bu = bursa, cx = coxa, fe = femur, op = operculum, po = postfemur, pre = prefemur, rdg = femoral ridge, st-pl = stigmatic plate, ta = tarsus, ti = tibia. Scale bars in millimetres.
Zephronia triforma sp. nov., ♂ paratype (MZKKU) — SEM. A–C. Overview of telopods (anterior, posterior, and ventral views, respectively); D. Crenulated teeth of posterior telopods, posterior view; E. Sclerotised spots on immovable finger of posterior telopod, sublateral view; F. Anterior telopod, ventral view; G. Inner horns, anterior view. Abbreviations: cr-t = crenulated teeth, ml = membranous lobe, scl-s = sclerotised spots. Scale bars in millimetres.
Endoterga of thoracic shields and midbody tergites — SEM. A, B. Zephronia dawydoffi Attems, 1953 (♂, MZKKU); C, D. Zephronia laotica Wesener, 2019 (♂, MZKKU); E, F. Zephronia tamarindina sp. nov. (♂ paratype, MZKKU); G, H. Zephronia triforma sp. nov. (♂ paratype, MZKKU). Abbreviations: cp = cuticular impression, ia = inner area, ma = middle area, oa = outer area, pm = posterior margin. Scale bars in millimetres.
Drawing of telopods of some Zephronia species. A–E. Zephronia dawydoffi Attems, 1953; F–J. Zephronia laotica Wesener, 2019; K–O. Zephronia tamarindina sp. nov.; P–T. Zephronia triforma sp. nov.; A, F, K, P. Sublateral view of left anterior telopods; B, G, L, Q. Anterior view of left anterior telopods; C, H, M, R. Posterior view of left anterior telopods; D, I, N, S. Anterior view of left posterior telopods; E, J, O, T. Posterior view of left posterior telopods. Abbreviations: cr-t = crenulated teeth, ml = membranous lobe, scl-s = sclerotised spots, syn-cx = syncoxite, sp = spine. Scale bars: 0.5 mm.
Distribution map of Zephronia species in Thailand. Number inside the circle indicates species: 1 = Z. chantaburiensis Srisonchai & Wesener, 2024; 2 = Z. chrysomallos Bhansali & Wesener, 2022; 3 = Z. dawydoffi Attems, 1953; 4 = Z. enghoffi Srisonchai & Likhitrakarn, 2021; 5 = Z. erawani Bhansali & Wesener, 2022; 6 = Z. golovatchi Srisonchai & Likhitrakarn, 2021; 7 = Z. lannaensis Likhitrakarn & Golovatch, 2021; 8 = Z. laotica Wesener, 2019; 9 = Z. macula Srisonchai & Wesener, 2024; 10 = Z. ovalis Gray, 1832; 11 = Z. panhai Srisonchai & Likhitrakarn, 2021; 12 = Z. phrain Likhitrakarn & Golovatch, 2021; 13 = Z. siamensis Hirst, 1907; 14 = Z. sukhothaiensis Srimongkol & Srisonchai, 2026; 15 = Z. tamarindina sp. nov.; 16 = Z. tratensis Srimongkol & Srisonchai, 2026; 17 = Z. triforma sp. nov.; 18 = Z. viridisoma Rosenmejer & Wesener, 2021. Colours (yellow, green, purple, red) indicate specimens examined in this study.
Measurements. Male holotype; body length = 17.9 mm; width (broadest), of thoracic shield = 10.6 mm, of tergite 7 = 11.1 mm; highest of thoracic shield = 5.8 mm, of tergite 7 = 6.5 mm. Males: body length = 15.2–26.2 mm; width (broadest), of thoracic shield = 8.7–11.03 mm, of tergite 7 = 9.2–15.2 mm; highest of thoracic shield = 5.2–8.7 mm, of tergite 7 = 5.3–9.7 mm. Females: body length = 18.2–30.8 mm; width (broadest), of thoracic shield = 8.9–15.2 mm, of tergite 7 = 9.6–15.9 mm; highest of thoracic shield = 5.4–9.2 mm, of tergite 7 = 5.1–11.1 mm.
Colouration (Fig.
Living specimens of Nakhon Sawan population with a brownish emerald-green body. Head and collum brownish emerald green. Thoracic shield and tergites dark green with dark brown posterior. Anal shield emerald green with pale brown on posterior margin. Antenna and legs green. Venter pale brown. Specimens in 95% ethanol faded to dark brown after 1.5 years of preservation.
Head (Fig.
Antennae (Fig.
Epipharynx (Fig.
Gnathochilarium (Fig.
Mandibles (gnathal lobe) (Fig.
Tegument (Figs
Collum (Figs
Thoracic shield. Anterior margin with shallow canal covered by numerous long setae. Phitsanulok population with 3–6 small keels. Nakhon Sawan population with 6–10 conspicuous keels.
Midbody tergite (Fig.
Anal shield (Fig.
Endotergum of thoracic shield and midbody tergite (Fig.
Pleurite (laterotergite) (Fig.
Subanal plate of female (Fig.
Stigmatic plates (Fig.
Legs (Fig.
Male sexual characters (Fig.
Anterior telopods (Figs
Posterior telopods (Figs
Female sexual characters (Fig.
Notable sexual dimorphism is observed in the Phitsanulok specimens, in which adult males possess a greyish brown body with dark brown freckles, whereas adult females exhibit a reddish-brown body lacking freckles. Specimens from Nakhon Sawan Province differ in body colouration (brownish emerald-green) from those of Phitsanulok Province. The two populations also show slight morphological variation in marginal bristles of endotergum, the Phitsanulok specimens having bristles reaching two-thirds the length of the outer area, while those from Nakhon Sawan reach only one-third. These differences are accompanied by moderate but sub-specific genetic differentiation between the two populations (pairwise distances 5.02–5.63%), which remains well below the lowest interspecific distance recovered here (8.68%). Specimens from the type locality (Ban Nam Kob in Phitsanulok Province) were heavily infested with numerous small, white, phoretic deutonymphs of an unidentified mite, concentrated on the ventral surface, particularly legs (Fig.
Zephronia triforma sp. nov. is currently known only from the upper central region of Thailand, specifically Nakhon Sawan and Phitsanulok Provinces (Fig.
The species name is a Latin compound noun derived from tri- (three) and forma (form), referring to the three distinct morphotypes observed in the material used to describe the new species.
An integrative approach combining both morphological and molecular information led to the discovery of two new giant pill-millipede species, viz., Z. tamarindina sp. nov. and Z. triforma sp. nov., and also the first records of Z. dawydoffi and Z. laotica in Thailand. All specimens of the latter two species closely match the original descriptions; however, morphological variations were observed and are documented here with SEM images. Both new species are assigned to the genus Zephronia based on the presence of diagnostic genus-specific characters, including axe-shaped antennae, the number of apical spines on the tarsi, and the distinctive morphology of the male telopods (
In Zephronia, species identification based solely on telopod morphology can be challenging, as interspecific variation in this structure is sometimes subtle and not always sufficient for clear species delimitation. Therefore, additional morphological characters, in combination with molecular evidence, are required to achieve reliable species identification.
Zephronia triforma sp. nov. is notable for its sexually dimorphic colouration within the Phitsanulok population, in which the males exhibit randomly distributed dark spots on a greyish-brown body, while females are reddish brown. This colour pattern differs markedly from that observed in both sexes of the Nakhon Sawan population, which display a uniform brownish emerald-green colouration and from all other known Zephronia species, with the exception of the spotted morph of Z. macula. Nevertheless, Z. triforma sp. nov. can be readily distinguished from Z. macula by several characters, including differences in live body colouration (Z. macula greenish to reddish brown vs. Z. triforma sp. nov. greyish brown), and clear genetic divergence (
In this study, DNA barcoding proved to be a highly effective tool for species delimitation (
In our study, the COI genetic distance between geographic populations of Z. triforma sp. nov. (5.02–5.63%) approaches, but remains below, the minimum interspecific distance observed between Z. siamensis and Z. dawydoffi (8.68%). High intraspecific sequence divergence is relatively common in giant pill-millipedes, likely driven by low dispersal ability and prolonged geographic isolation (
The relatively large interspecific genetic distances (8.68–24.89%) between the two newly described species, the two newly recorded species from Thailand, and their congeners (Sphaerobelum spp., Prionobelum spp., Cryxus, and Zephronia spp. from Southeast Asia) nonetheless highlight the evolutionary and biogeographical processes that have shaped diversity of Thai giant pill-millipedes (
Molecular data based on the COI barcoding region show that traditional generic boundaries within Zephroniinae are problematic. Specifically, Zephronia is non-monophyletic, with species like Z. hui, Z. medogensis, and Z. zhouae nesting within Sphaerobelum. Together with the close placement of Prionobelum and Cryxus, these trees show that traditional diagnostic traits such as telopod morphology and the position of the Tömösváry's organ have evolved independently across different lineages or show greater structural variation than previously recognised. While single-locus COI data cannot formally revise the subfamily classification, they clearly demonstrate that morphology-based generic limits require thorough reconsideration. To establish a stable classification for Zephroniinae, future studies incorporating multi-locus markers, more conserved nuclear regions, and genomic-scale data combined with more comprehensive taxon sampling across Southeast Asia could improve the resolution within the genus or family (
Based on the distribution record obtained in this study, Z. tamarindina sp. nov. and Z. triforma sp. nov. appear to be restricted to the upper central region of Thailand. Z. tamarindina sp. nov. is currently known only from its type locality in Phetchabun Province, whereas Z. triforma sp. nov. has been recorded exclusively from Nakhon Sawan and Phitsanulok Provinces. Consequently, both species are considered endemic to the upper central region. Z. tamarindina sp. nov. exhibits a narrow distribution, similar to several other Thai species that are confined to their type localities, such as Z. chantaburiensis, Z. chrysomallos, Z. erawani, and Z. viridisoma (Fig.
Zephronia dawydoffi, originally described from southern Cambodia (Ream, Sre Umbell; Kampot, Kampong Speu) (
Zephronia laotica was originally reported from Champasak Province in southern Laos, on the east bank of the Mekong River, and in this study it was recorded from Si Sa Ket Province in northeastern Thailand, approximately 150–200 km from its type locality (Fig.
This study increases the number of recognised Zephronia species in Thailand to 18, expanding the genus’s known range, particularly in northeastern and upper central Thailand. The steady discovery of new giant pill-millipede species since 2021 (e.g.,
This work was conducted as part of a series of small workshops on soil fauna surveys in the Chao Phraya and Mekong River basins, organised for undergraduate and graduate students in the Young Taxonomist program at Khon Kaen University. We are grateful to the curators of NHMW (Nesrine Akkari) and ZFMK (Thomas Wesener) for providing information on the type material for comparison with the newly collected specimens. We sincerely thank the Department of Biology, Faculty of Science, Khon Kaen University, for providing laboratory facilities throughout this study. We also thank the field assistants, students from Khon Kaen University Museum of Zoology (MZKKU) and local partners for their assistance with specimen collection and site access. We thank the staff of the Department of National Parks, Wildlife and Plant Conservation for their guidance during field surveys on soil fauna in the Chao Phraya River Basin areas. Special thanks are due to Khanet Srikampha for providing photographs of live specimens. We also acknowledge the constructive comments from reviewers and journal editors, which improved the quality of this work. Computational analyses were supported by the Cipres Science Gateway. Nisarat Tungpairojwong and Ruttapon Srisonchai jointly supervised this study and shared responsibility for the preparation, revision, and correspondence of the manuscript; Nisarat Tungpairojwong is listed as the single corresponding author in accordance with the journal’s submission requirements.
The authors have declared that no competing interests exist.
The animal ethic protocol was approved by the Institutional Animal Care and Use Committee of Khon Kaen University (No. IACUC-KKU-137/67).
The authors accept full responsibility for the content of the manuscript, including the disclosure of any use of AI.
No AI tools were used in the preparation of this manuscript.
This work was supported by the Fundamental Fund, Fiscal year 2024 (200596).
Conceptualization: RS, NT; Methodology: CW, RS, WK, NT; Validation: WK, NT; Formal analysis: CW, RS; Investigation: CW, RS, NT; Resources: RS, NT; Data curation: CW; Writing-Original Draft: CW; Writing - Review & Editing: RS, WK, NT; Visualization: CW; Supervision: RS, NT; Project administration: RS, NT; Funding acquisition: NT.
C. Wilaisorn https://orcid.org/0009-0001-3673-4167
R. Srisonchai https://orcid.org/0000-0002-7142-0999
W. Kutanan https://orcid.org/0000-0001-7767-1644
N. Tungpairojwong https://orcid.org/0000-0001-8135-4246
The data underlying this article are available in the GenBank Nucleotide Database (https://www.ncbi.nlm.nih.gov/genbank/), and can be accessed with accession numbers listed in Table
Supplementary information
Data type: xlsx
Explanation note: Genetic divergences of cytochrome c oxidase subunit I (COI) sequence (uncorrected p-distances) within and among Zephronia species, and related genera (Sphaerobelum, Prionobelum, and Cryxus).