Research Article |
Corresponding author: Aiki Yamada ( aiki.ymd@gmail.com ) Academic editor: Matthew Prebus
© 2023 Aiki Yamada, Dai Dac Nguyen, Katsuyuki Eguchi.
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:
Yamada A, Nguyen DD, Eguchi K (2023) First discovery of the ant genus Eburopone Borowiec, 2016 (Hymenoptera, Formicidae, Dorylinae) in the Oriental realm, with description of a new species from Vietnam. ZooKeys 1184: 1-17. https://doi.org/10.3897/zookeys.1184.109702
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The doryline ant genus Eburopone Borowiec, 2016 currently contains only one valid species, E. wroughtoni (Forel, 1910) from southern Africa, with a considerable number of undescribed species awaiting formal description in the Afrotropical and Malagasy regions. In the present paper, Eburopone easoana sp. nov. is described based on workers and dealate queens from a colony series collected in an evergreen forest on the Dak Lak Plateau of Vietnam (Ea So Nature Reserve, Dak Lak Province). The worker of the new species is morphologically clearly distinguished from E. wroughtoni by the combination of following characteristics: i) frontal line distinct, extending a little beyond mid-length of cranium; ii) anterior (frontoclypeal) margins of torulo-posttorular complex not forming conspicuous lobes protruding over anterior clypeal margin in full-face view; iii) mandibles when closed in full-face view forming only a little space between anterior clypeal margin and mandibles; iv) promesonotal suture faint and inconspicuous; v) abdominal segment III in dorsal view distinctly wider than long, with lateral margins only feebly convex. This represents the first discovery of the genus Eburopone in the Oriental realm, revealing the disjunct distribution of the genus. A partial sequence of the mitochondrial COI gene (658 bp) is provided as a DNA barcode for the new species. A worker-based key to the doryline genera of the Oriental realm is also provided.
Central Highlands, disjunct distribution, Indochina, morphology, non-army ant dorylines, taxonomy, Tay Nguyen
The subfamily Dorylinae (Hymenoptera: Formicidae) is a clade of predatory ants primarily occurring in the tropics and subtropics of the world, consisting of true army ants and their relatives (
Eburopone Borowiec, 2016 was one of the two genera newly established by the reclassification of the former Cerapachys members in
Eburopone wroughtoni was originally described as Cerapachys wroughtoni based on workers from South Africa. Forel later described E. w. var. rhodesiana (Forel, 1913) and E. roberti (Forel, 1914) from Zimbabwe and South Africa, respectively. The latter two names were synonymized under C. wroughtoni by Brown (1975), for which the genus Eburopone was established by
During our recent fieldwork in the Ea So Nature Reserve, Dak Lak Province, Central Highlands (Tay Nguyen) of Vietnam, we collected a colony fragment of an unknown doryline species. After careful morphological examination, this species is found to fit with the concept of Eburopone but is well distinguished from the only valid species E. wroughtoni. This represents the first discovery of the genus from the Oriental realm and reveals the disjunct distribution of the genus (Fig.
Abbreviations of the specimen depositories are as follows:
IEBR Institute of Ecology and Biological Resources, Vietnam Academy of Science and Technology, Hanoi, Vietnam;
General morphological observations were made under epi-illumination using a Nikon SMZ1270 stereo microscope (Nikon, Tokyo, Japan) and a Nikon AZ100 multi-zoom microscope. Focus-stacked images of specimens under epi-illumination were produced by Affinity Photo 2.1.0 (Serif (Europe), Nottingham, UK) based on source images taken using a Nikon Z50 digital camera attached to the latter microscope via a NY-1S camera adapter (Micronet, Saitama, Japan). The retouching and adjusting functions of the software were used to improve the quality and clarity of stacked images.
Two workers from the same colony as the type series were used for further observations of some body parts by scanning electron microscope (SEM) JEOL JSM-6510 (JEOL, Tokyo, Japan) and transmission light microscope (TLM) Nikon Eclipse E600 (one of the specimens was also used for DNA extraction before conducting SEM and TLM observations). SEM was used to observe some morphological details of the cranium, promesonotal suture, and pronotomesopleural junction: the focal body parts were dissected and coated with platinum using a spatter coater JEOL JFC-1600 before SEM observation. TLM was used to observe the mouthparts: the mouthparts were dissected after cleaning by the DNA extraction protocol with Proteinase K described below, and slide-mounted with Euparal. The partly focus stacked images of the mouthparts under TLM were produced using Helicon Focus Pro 8.2.2 (Helicon Soft, Kharkov, Ukraine) based on source images taken by a Nikon Z5 digital camera attached to the TLM via a NY-1S35 camera adapter (Micronet).
Fifteen (or sixteen for queens) linear morphometric characters were measured using ImageJ 1.53e (National Institutes of Health, USA, available at https://imagej.nih.gov/ij/) based on photographs taken using a Nikon Z5 digital camera attached to a Nikon SMZ1270 stereo microscope via a NY-1S35 camera adapter (Micronet), and then ten (or eleven for queens) indices were calculated. Definitions of the worker morphometric characters basically follow those in
HL Head Length: maximum length of cranium in full-face view, measured from the midpoint of the anterior clypeal margin to the midpoint of the posterior margin of cranium;
HW Head Width: maximum width of cranium in full-face view (excluding compound eyes);
EL Eye Length: maximum diameter of compound eye in lateral view (for queens);
OL Ocellus Length: maximum diameter of median ocellus (for queens);
SL Scape Length: maximum length of antennal scape excluding basal condylar bulb;
WL Weber’s Length of Mesosoma: maximum diagonal distance of mesosoma in lateral view, measured from the angle at which the pronotum meets the cervix to posteroventral angle of metapleuron;
DML Dorsal Mesosomal Length: length of mesosomal dorsum from midpoint of anterodorsal margin of pronotum to midpoint of dorsal margin of propodeal declivity in dorsal view;
PH Pronotal Height: maximum height of pronotum in lateral view (for workers);
PW Pronotal Width: maximum width of pronotum in dorsal view;
MFL Metafemur Length: maximum length of metafemur, measured in dorsal view;
PTH Petiolar Height: maximum height of the petiolar tergum (abdominal tergum II) in lateral view, excluding petiolar sternum;
PTL Petiolar Length: maximum length of petiole (abdominal segment II) in dorsal view, measured from anterodorsal corner to posterior margin of posterior petiolar peduncle;
PTW Petiolar Width: maximum width of petiole (abdominal segment II) in dorsal view;
A3L Abdominal Segment III Length: maximum length of abdominal segment III in dorsal view, excluding presclerites (helcium);
A3W Abdominal Segment III Width: maximum width of abdominal segment III in dorsal view;
A4L Abdominal Segment IV Length: maximum length of abdominal segment IV in dorsal view, excluding presclerites;
A4W Abdominal Segment IV Width: maximum width of abdominal segment IV in dorsal view;
CI Cephalic Index: HW / HL × 100;
SI Scape Index: SL / HL × 100;
EI Eye Index: EL / HL × 100 (for queens);
OI Ocellus Index: OL / HL × 100 (for queens);
DMI Dorsal Mesosoma Index: PW / WL × 100;
DMI2 Dorsal Mesosoma Index 2: DML / WL × 100;
LMI Lateral Mesosoma Index: PH / WL × 100 (for workers);
MFI Metafemur Index: MFL / HW × 100;
LPI Lateral Petiole Index: PTL / PTH × 100;
DPI Dorsal Petiole Index: PTW / PTL × 100;
DA3I Dorsal Abdominal Segment III Index: A3W / A3L × 100;
DA4I Dorsal Abdominal Segment IV Index: A4W / A4L × 100.
The morphological terminology follows
A partial sequence of mitochondrial cytochrome c oxidase subunit I gene (COI, 658 bp of standard DNA barcoding region) was determined from a single worker from the same colony as the type series. Total DNA was extracted using the Chelex-TE-ProK protocol: the specimen was washed and dissected in TE buffer, incubated at 56 °C for 24 h with 105 μL of the extraction buffer which contained 100 μL of 10% Chelex-TE solution (Chelex-100 resin, 143-2832, Bio-Rad Laboratories, California, USA) and 5 μL of Proteinase K (QIAGEN, Venlo, Netherlands), and lastly heated at 99 °C for 10 min for inactivating Proteinase K in the extraction buffer. PCR was performed with the primers LCO-EG (TTTCAACAAATCACAAAGAYATYGG) and HCO-EG (TAAACTTCAGGRTGACCRAAAAATCA). The PCR contained 5 μL of KOD One PCR Master Mix (KMM-101, TOYOBO, Osaka, Japan), 3.9 μl of distilled water, 0.3 μL of 10 pmol/μL forward and reverse primers (final 0.3 μM), and 1 μL of the DNA extract. The PCR thermal cycling condition was 40 cycles of 10 s at 98 °C, 5 s at 48.5 °C, and 5 s at 68 °C. After confirming the PCR amplification on a 2.0% agarose gel, the amplified product was incubated at 37 °C for 4 min and 80 °C for 1 min with ExoSAP-IT Express (Applied Biosystems, California, USA) to remove any excess primers and nucleotides. The cycle sequencing reaction was run with SupreDye Cycle Sequencing Kit v. 3.1 (AdvancedSeq, California, USA). The sequencing reaction product was purified and concentrated by ethanol precipitation with sodium acetate, and the nucleotide sequence was determined using an automated sequencer 3730xl DNA Analyzer (Applied Biosystems). Sequence assembly was performed using ChromasPro 1.7.6 (Technelysium, Queensland, Australia).
Eburopone Borowiec, 2016: 124. Type-species by original designation and monotypy: Cerapachys wroughtoni Forel, 1910.
For general morphological diagnosis and description of the genus, see
Holotype. Vietnam • worker; Vietnam, Dak Lak Province, Ea Kar District, Ea So Nature Reserve; 12.9676°N, 108.5230°E, 478 m alt.; 17 Sept. 2019; K. Eguchi leg.; colony code Eg17ix19-297; IEBR.
Paratypes. 6 workers and 2 dealate queens from the same colony as the holotype; IEBR,
Two workers from the same colony as the type series, used for SEM and dissecting observations (one of these was used also for DNA barcoding).
A partial mitochondrial COI sequence of 658 bp length determined from a non-type worker (the same colony as the type series) is deposited at GenBank: accession number, LC776907.
Body rather bicolored: abdominal segments III–VII distinctively paler-colored than most surfaces of cranium and mesosoma. Frontal line distinct, extending a little beyond mid-length of cranium. Occipital corner in lateral view strongly produced posteriad to form conspicuous angle. Anterior (frontoclypeal) margins of torulo-posttorular complex not forming conspicuous lobes protruding over anterior clypeal margin in full-face view. Mandibles when closed in full-face view forming only a little space between anterior clypeal margin and mandibles. Promesonotal suture faint and inconspicuous. Petiole in dorsal view almost as long as or slightly longer than wide, with weakly convex lateral margins. Subpetiolar process in lateral view rounded lobate, with anterobasal margin weakly emarginate; posteroventral slope gentle, weakly concave. Abdominal segment III in dorsal view subtrapezoidal, strongly wider posteriorly, distinctly wider than long.
Holotype : HL 0.63, HW 0.55, SL 0.31, WL 0.86, DML 0.73, PH 0.31, PW 0.37, MFL 0.43, PTH 0.26, PTL 0.29, PTW 0.28, A3L 0.35, A3W 0.46, A4L 0.58, A4W 0.62, CI 87, SI 50, DMI 43, DMI2 85, LMI 36, MFI 79, LPI 111, DPI 97, DA3I 131, DA4I 107.
Paratypes (N = 6): HL 0.63–0.65, HW 0.55–0.57, SL 0.31–0.33, WL 0.89–0.92, DML 0.74–0.80, PH 0.32–0.35, PW 0.37–0.40, MFL 0.44–0.47, PTH 0.27–0.29, PTL 0.30–0.33, PTW 0.29–0.32, A3L 0.37–0.43, A3W 0.47–0.51, A4L 0.56–0.64, A4W 0.64–0.66, CI 86–88, SI 48–51, DMI 42–43, DMI2 84–87, LMI 36–38, MFI 80–83, LPI 111–114, DPI 95–98, DA3I 118–130, DA4I 103–112.
Paratypes (N = 2): HL 0.70–0.72, HW 0.58–0.61, EL 0.19–0.20, OL 0.05–0.06, SL 0.35–0.36, WL 1.18–1.20, DML 1.07–1.08, PW 0.57–0.58, MFL 0.51–0.53, PTH 0.31, PTL 0.35–0.36, PTW 0.35–0.37, A3L 0.48–0.49, A3W 0.56–0.58, A4L 0.82, A4W 0.73–0.75, CI 84, SI 49–50, EI 28, OI 7–8, DMI 48, DMI2 90–91, MFI 87–88, LPI 111–117, DPI 100–101, DA3I 115–119, DA4I 89–91.
Worker. Body coloration. Body rather bicolored: cranium, mesosoma, and petiole, mostly dark reddish brown; antennae, anterior part of cranium, mandibles, legs, and abdominal segments III–VII paler brownish to yellowish (Fig.
Head structure. Cranium in full-face view subrectangular, 1.14–1.17× longer than wide (CI, 86–88), with convex lateral margins; posterior margin widely weakly concave. Occipital corner in lateral view strongly produced posteriad to form conspicuous angle. Occipital carina (oc, Fig.
Cranium morphology of Eburopone easoana sp. nov. workers, colony Eg17ix19-297, holotype (A–C, E) and nontype (D) A head in lateral view B occipital corners in posterior view C anterior part of cranium in ventrofrontal view D scanning electron microscopic (SEM) photograph of anterior part of cranium in dorsofrontal view, antennae and mandibles removed (left antennal bulbus remains) E torulo-posttorular complex and parafrontal ridges in posterior vertical view against its posterior face. Black arrows in A and C indicate a weak protrusion on lateroclypeal teeth. Abbreviations: ala – atalar acetabulum; oc – occipital carina; fl – frontal line; lct – lateroclypeal teeth; pfr – parafrontal ridge; tptc – torulo-posttorular complex.
Cranial venter and mouthparts morphology of Eburopone easoana sp. nov. workers, colony Eg17ix19-297, nontypes A scanning electron microscopic (SEM) photograph of cranium in ventral view, mandibles removed B transmission light microscopic (TLM) photograph of labrum in frontal view, partly focus-stacked to emphasize lateral labral processes on caudal surface C TLM photograph of right maxilla in ventral view, with maxillary palpomeres indicated by black arrows D TLM photograph of labium in oblique lateral view, left side, with labial palpomeres indicated by black arrows. Abbreviations: oc – occipital carina; ala – atalar acetabulum; hyt – hypostomal teeth; lbrp – lateral labral process; pgr – postgenal ridge.
Mesosomal structure.
Mesosoma with evenly and slightly convex dorsum in lateral view (Fig.
Mesosoma and abdominal segments II–III morphology of Eburopone easoana sp. nov. workers, colony Eg17ix19-297, holotype (A, B) and non-type (C, D) A mesosoma and abdominal segment II–III in lateral view B mesosoma and abdominal segment II–III in dorsal view C scanning electron microscopic (SEM) photograph of pronotomesopleural junction in lateral view D SEM photograph of promesonotum in dorsal view. Abbreviations: epp – pleural endophragmal pit; pmns – promesonotal suture.
Metasomal structure. Petiole (abdominal segment II) in dorsal view 1.02–1.05× longer than wide (length measured from anterodorsal corner to posterior margin of posterior petiolar peduncle; DPI, 95–98), with weakly convex lateral margins; posterior margin of posterior petiolar peduncle slightly arched anteriad (Fig.
Abdominal segments IV–VII morphology of Eburopone easoana sp. nov. workers, colony Eg17ix19-297, holotype A abdominal segments IV–VII in dorsal view B abdominal segments IV–VII in ventral view, with the putative glandular patch of poststernite IV indicated by a black arrow C cinctus and presclerites of abdominal segment IV in dorsal view D abdominal tergite VII (pygidium) in dorsoposterior view.
Setation and sculpture. Body setation relatively sparse. Anterolateral surfaces of cranium coarsely longitudinally rugose (Fig.
Dealate queen. Largely similar to the worker except for some queen-specific features. Eyes and ocelli large and conspicuous (Fig.
Habitus of dealate queens of Eburopone easoana sp. nov., colony Eg17ix19-297, paratypes (all except D are of the same individual) A head in full-face view B habitus in lateral view C habitus in dorsal view D abdominal segment IV–VII in ventral view, with the putative glandular patch of poststernite IV indicated by a black arrow.
Male. Unknown.
The specific epithet is named after the type locality, Ea So Nature Reserve: easo combined with the Latin feminine suffix -ana, adjective.
The collecting site of the type colony series is primarily covered with a relatively disturbed secondary evergreen forest. The collector (K. Eguchi) did not record the exact microhabitat and collecting situation of the colony fragment.
The worker of E. easoana is morphologically easily distinguished from the only other valid congener E. wroughtoni from southern Africa (see also the account of E. wroughtoni below) by the combination of following characteristics: i) frontal line distinct, extending a little beyond mid-length of cranium; ii) anterior (frontoclypeal) margins of torulo-posttorular complex not forming conspicuous lobes protruding over anterior clypeal margin in full-face view (anterior clypeal margin evenly weakly concave in full-face view); iii) mandibles when closed in full-face view forming only a little space between anterior clypeal margin and mandibles (basal angles nearly reaching center line of cranium when mandibles closed); iv) promesonotal suture faint and inconspicuous; v) abdominal segment III in dorsal view distinctly wider than long, with lateral margins only feebly convex.
The morphology of workers of E. easoana is generally consistent with the concept of Eburopone in
Cerapachys wroughtoni
Forel, 1910: 422.
Cerapachys wroughtoni var. rhodesiana
Forel, 1913: 212.
Cerapachys roberti
Forel, 1914: 212.
The following type materials of E. wroughtoni and its two junior synonyms were non-physically examined based on the images available from AntWeb (https://www.antweb.org): E. wroughtoni (Forel, 1910), syntype, CASENT0249876 (
Although Brown (1975) treated two southern African Eburopone (sub)species, E. roberti and E. w. rhodesiana as junior synonyms of E. wroughtoni, the types of these names show conspicuous morphological differences, especially between E. w. rhodesiana and the others. Eburopone w. rhodesiana is the most morphologically distinct among these by i) lateroventral flange of occipital carina less developed; ii) promesonotal suture inconspicuous (hardly recognizable in the examined image); iii) abdominal segment III distinctly wider than long. Eburopone roberti is different from the syntype of E. wroughtoni at least by having deeper and more prominent promesonotal suture. The differences suggest a possible heterospecificity of these names, especially for E. w. rhodesiana. However, their taxonomic statuses are not addressed in the present study, since it would be appropriate to examine additional colony series; additionally, many undescribed species are known from the Afrotropical and Malagasy regions. Even if multiple species are confused under E. wroughtoni, the independence of E. easoana at the species level is unquestionable.
The following key is based on the generic descriptions and global key to genera of Dorylinae by
1 | Abdominal tergite VII (last visible tergite, pygidium), not armed with numerous differentiated stout chaetae (“peg-like setae”), at most with only a few pairs of cuticular projections | 2 |
– | Abdominal tergite VII armed with numerous stout chaetae | 3 |
2 | Abdominal segment III forms differentiated “postpetiole”, narrowly attached to segment IV | Aenictus Shuckard, 1840 |
– | Abdominal segment III not differentiated and broadly attached to segment IV | Dorylus Fabricius, 1793 |
3 | Abdominal segment III broadly attached to segment IV, without a conspicuous anterior constriction of segment IV | Yunodorylus Xu, 2000 |
– | Abdominal segment III at least weakly differentiated from segment IV, with a conspicuous anterior constriction of segment IV | 4 |
4 | Meso- and metatibiae each with two spurs; most body surface with prominent costate sculpture | Chrysapace Crawley, 1924 |
– | Mesotibiae with a single spur or without spurs and metatibiae always with a single spur | 5 |
5 | Mesotibiae without spurs | Simopone Forel, 1891 |
– | Mesotibiae with a single spur | 6 |
6 | Petiole (abdominal segment II) dorsolaterally marginate at least in anterior portion; metacoxae usually with a conspicuously lamellate posterior flange | Lioponera Mayr, 1879 |
– | Petiole (abdominal segment II) not conspicuously dorsolaterally marginate; if petiole appearing marginate, metacoxae without a conspicuously lamellate posterior flange | 7 |
7 | Pronotum and mesopleuron completely fused, never with a deep cut of pronotomesopleural junction | 8 |
– | Pronotum and mesopleuron unfused, with a deep cut of pronotomesopleural junction | 9 |
8 | Constrictions present at anterior end of abdominal segments V and VI | Zasphinctus Wheeler, 1918 |
– | Constrictions absent at anterior end of abdominal segments V and VI | Parasyscia Emery, 1882 |
9 | Constrictions present at anterior end of abdominal segments V and VI | Eusphinctus Emery, 1893 |
– | Constrictions absent at anterior end of abdominal segments V and VI | 10 |
10 | Antennae 12-merous | 11 |
– | Antennae 8- to 11-merous | 12 |
11 | Eyes conspicuous with more than 20 ommatidia; abdominal sternite IV normal without a whitish patch near the posterior edge | Cerapachys Smith, 1857 |
– | Eyes absent or perhaps vestigial with at most several weakly differentiated ommatidia; abdominal sternite IV with a unique whitish patch near the posterior edge (may be feeble) | Eburopone Borowiec, 2016 |
12 | Abdominal tergite IV in lateral view folding over sternite IV and the anterior portion of sternite at least partly obscured; abdominal segment III relatively wide in dorsal view and larger than the preceding abdominal segment II (petiole) | Syscia Roger, 1861 |
– | Abdominal tergite IV not folding over sternite IV and the anterior portion of the sternite visible; abdominal segment III relatively narrow in dorsal view and similar in size to the preceding abdominal segment II (petiole) | Ooceraea Roger, 1862 |
We would like to thank Drs Nguyen Van Sinh (Director of the Institute of Ecology and Biological Resources (IEBR), Vietnam Academy of Science and Technology, Vietnam), Truong Xuan Lam (Vice Director of IEBR), Nguyen Duc Anh (IEBR), and staff of Ea So Nature Reserve (Dak Lak Province, Vietnam) for their help in the field survey and managing research permissions; Drs Marek L. Borowiec and Mamoru Terayama for their reviews and comments on the manuscript. The present study was conducted under the framework of the memorandum of understanding (MOU) on academic research cooperation between Tokyo Metropolitan University and IEBR.
The authors have declared that no competing interests exist.
No ethical statement was reported.
This study has been supported by the following funds: the Fund for the Promotion of Joint International Research (Fostering Joint International Research (B)) (JSPS KAKENHI, No. 22KK0087, Leader: K. Eguchi, FY2022–2025), the Tokyo Metropolitan University Fund for TMU Strategic Research (Leader: Prof. Noriaki Murakami, FY2020–FY2022), and the Asahi Glass Foundation (Leader: K. Eguchi, FY2017–FY2022).
AY conducted morphological examinations, wrote most of the main text including descriptions, took specimen images and prepared all figures. DN and KE conducted field survey to collect specimens, also contributed writing and editing on the manuscript.
Aiki Yamada https://orcid.org/0000-0002-2141-0661
Dai Dac Nguyen https://orcid.org/0009-0006-8024-2553
Katsuyuki Eguchi https://orcid.org/0000-0002-1054-1295
All of the data that support the findings of this study are available in the main text.