Zootaxa 5374 (4): 505–518 ISSN 1175-5326 (print edition)

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Copyright © 2023 Magnolia Press
Article ZOOTAXA
ISSN 1175-5334 (online edition)
https://doi.org/10.11646/zootaxa.5374.4.3
http://zoobank.org/urn:lsid:zoobank.org:pub:5DA74ABE-380E-4CDE-A0F3-5A9A4A2881C8

A new species of Vietnamophryne (Anura: Microhylidae) from Northeastern

Vietnam
HOA THI NINH1, LINH TU HOANG LE1, HAI TUAN BUI1, HUY QUOC NGUYEN2, NIKOLAI ORLOV3,
OLGA BEZMAN MOSEYKO3, MANH VAN LE4, SANG NGOC NGUYEN4, CHUNG VAN HOANG5, THOMAS
ZIEGLER6,7 & TAO THIEN NGUYEN1,*
1
Institute of Genome Research, Vietnam Academy of Science and Technology, 18 Hoang Quoc Viet Road, Hanoi, Vietnam
� ninhhoa.hnue@gmail.com; https://orcid.org/0009-0002-8133-432X
� letuhoanglinh98@gmail.com; https://orcid.org/0009-0003-0887-2786
� tuanhai@eulipotyphla.com; https://orcid.org/0000-0003-4065-7229
2
Vietnam National Museum of Nature, Vietnam Academy of Science and Technology, 18 Hoang Quoc Viet Road, Hanoi, Vietnam
� huynguyen17295@gmail.com; https://orcid.org/0000-0003-3171-1561
3
Zoological Institute, Russian Academy of Sciences, Universitetskaya nab. 1, St. Petersburg, 199034, Russia
� azemiops@zin.ru; https://orcid.org/0000-0003-4401-348X
� olga.bezman-moseyko@zin.ru; https://orcid.org/0000-0002-5564-4415
4
Institute of Tropical Biology, Vietnam Academy of Science and Technology; 9/621 Hanoi Highway, Ho Chi Minh City, Vietnam
� leemanh94@gmail.com; https://orcid.org/0000-0002-7401-4830
� ngocsangitb@yahoo.com; https://orcid.org/0000-0003-1955-3329
5
Forest Resources and Environment Centre, 300 Ngoc Hoi Road, Thanh Tri, Hanoi, Vietnam
� chungiebr@gmail.com; https://orcid.org/0000-0002-0709-974X
6
AG Zoologischer Garten Köln, Riehler Strasse 173, D-50735 Cologne, Germany
7
Institute of Zoology, University of Cologne, Zülpicher Straße 47b, D-50674 Cologne, Germany
� ziegler@koelnerzoo.de; https://orcid.org/0000-0002-4797-609X
*
Corresponding author: � nguyenthientao@gmail.com; https://orcid.org/0000-0002-5640-4536

Abstract

A new species of the genus Vietnamophryne is described from Vietnam on the basis of two specimens collected from
Tuyen Quang Province, Northeastern Vietnam. The new species is morphologically most similar to Vietnamophryne
occidentalis from Thailand, however, it differs from the latter by having large black blotches in the lower jaw region,
and a yellow-orange chest and belly. The genetic distance between the new species and other Vietnamophryne taxa is >
2.13% (16S mtDNA gene fragment). Vietnamophryne aurantifusca sp. nov. can be distinguished from all other species
of Vietnamophryne by a combination of the following morphological characteristics: Size medium (SVL 17.6–18.2 mm
in males); head wider than long; tympanum medium; finger I longer than half of finger II; dorsal skin relatively smooth
with some round nodules, concentrated in the middle of the back, arranged along the length of the back, with a prominent
ridge along the spine; Dorsum orangish-brown entirely and paler on margin of back with a small brownish ridge along the
spine; sides brownish with creamy patches and orange spots; ventral surface orange, with grey marbling, most intense on
the throat, ventral side of arms and thighs, and ventral surfaces of limbs dark grey with some orange spots.

Key words: Genetic distance, morphology, Vietnamophryne aurantifusca sp. nov., taxonomy, Tuyen Quang Province

Introduction

Northeastern Vietnam is located in a subtropical region, characterized by a warm climate, high rainfall and complex
topography. The region has a typical karst landscape with evergreen forests (Sterling et al. 2006). Although a surge
of field surveys on biodiversity has been carried out in this area, the amphibian fauna is still imperfectly studied.
Some new species of amphibians have been discovered from this area in the past five years, namely Zhangixalus
jodiae Nguyen, Ninh, Orlov, Nguyen, and Ziegler, 2020; Z. franki Ninh, Nguyen, Orlov, Nguyen, and Ziegler, 2020;
and Theloderma khoii Ninh, Nguyen, Nguyen, Hoang, Siliyavong, Nguyen, Le, Le, and Ziegler, 2022 (Nguyen et
al. 2020, Ninh et al. 2020, 2022).

Accepted by E. Quah: 19 Oct. 2023; published: 20 Nov. 2023 505

 The genus Vietnamophryne currently contains five species, and four of them are known only from Vietnam
at the current time, viz. Vietnamophryne orlovi Poyarkov, Suwannapoom, Pawangkhanant, Aksornneam, Duong,
Korost, and Che, 2018 from Phia Oac-Phia Den National Park, V. cuongi Nguyen, Hoang, Jiang, Orlov, Ninh,
Nguyen, Nguyen, and Ziegler, 2021 from Ba Vi National Park, V. vuquangensis Hoang, Jiang, Nguyen, Orlov, Le,
Nguyen, Nguyen, Nguyen, Nguyen, and Ziegler, 2021from Vu Quang National Park, and V. inexpectata Poyarkov,
Suwannapoom, Pawangkhanant, Aksornneam, Duong, Korost, and Che, 2018 from Kon Chu Rang Nature Reserve.
During our recent fieldwork in Northeastern Vietnam, a small microhylid species was collected from Sinh Long
Commune, Na Hang District, Tuyen Quang Province. Based on molecular and morphological data it proved to be a
representative of the genus Vietnamophryne.
At first glance, the specimens from Tuyen Quang resemble Vietnamophryne inexpectata (Poyarkov et al. 2018).
However, closer morphological examination revealed these taxa are morphologically distinct from each other. The
phylogenetic analyses showed that the new species is rather nested in a clade with Vietnamophryne cuongi, V. occidentalis
Poyarkov, Suwannapoom, Pawangkhanant, Aksornneam, Duong, Korost, and Che, 2018 and V. orlovi with high support
values. Thus, the microhylid frogs from Tuyen Quang Province represent a distinct species with genetic distances of
2.13–6.5% from its congeners. We herein describe this recently discovered population as a new species.

Material and methods

Sampling

A field survey was conducted in Sinh Long Commune, Na Hang District, Tuyen Quang Province, Vietnam from
20 to 30 May 2023 by Nguyen H.Q., Nguyen S.N., Le M.V., Le, L.T.H., Olga B.M., (Hereafter Nguyen et al.).
Geographic coordinates and elevation were obtained using a Garmin GPSMAP 78S (WGS 84 data). All specimens
were photographed in life prior to anesthetization with ethyl acetate and subsequent fixation in 75% ethanol
before finally being stored in 70% ethanol. DNA samples for molecular analyses were taken from the liver prior to
preservation and stored in 95% ethanol. Sex was determined by gonadal dissection. The type series was subsequently
deposited in the Institute of Ecology and Biological Resources (IEBR), Hanoi, Vietnam.

Molecular analyses

Extraction of genomic DNA from two samples (Table 1) was carried out using DNeasy Blood & Tissue Kits (QIAGEN,
Germany), the protocol followed the manufacturers’ instructions. Total DNA was amplified using an Eppendorf PCR
machine. PCR total volume was 20μl, consisting of 10μl of Mastermix, 7μl of water, 0.5μl of each primer at a concentration
of 10 pmol/μl, and 2 μl of DNA. Primers used in the PCR and for sequencing a fragment of 16S rRNA were as follows:
tVal-L: (5’-CGTACCTTTTGCATCATGGTC-3’, forward), H3056: (5’-CTCCGGTCTGAACTCAGATCACGTAGG-
3’, reverse) (Hedges 1994). PCR conditions: 94°C for 5 minutes of initial denaturation; with 30 cycles of denaturation
at 94°C for 30s, annealing at 56°C for 30s, and extension at 72°C for 45s; and the final extension at 72°C for 7 minutes.
PCR products were sent to the 1st Base company in Malaysia for sequencing. The obtained sequences were deposited
in GenBank under the accession numbers as stated in Table 1.

TABLE 1. Samples of Vietnamophryne and other species used for molecular analysis in this study.
No. Scientific name Voucher Locality Genbank Acc. Source
No.
1 Vietnamophryne IEBR A.5242 Tuyen Quang, Vietnam LC782740 This study
aurantifusca sp. nov.
2 Vietnamophryne IEBR A.5243 Tuyen Quang, Vietnam LC782741 This study
aurantifusca sp. nov.
3 V. cuongi VNMN10357 Ba Vi NP, Hanoi, Vietnam MT819964 Nguyen et al. (2021)
4 V. cuongi VNMN10358 Ba Vi NP, Hanoi, Vietnam MT819965 Nguyen et al. (2021)
...Continued on the next page

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 TABLE 1. (Continued)
No. Scientific name Voucher Locality Genbank Acc. Source
No.
5 V. cuongi VNMN10359 Ba Vi NP, Hanoi, Vietnam MT819966 Nguyen et al. (2021)
6 V. inexpectata ZMMU A5820 Kon Chu Rang NR, Gia MH004403 Poyarkov et al. (2018)
Lai, Vietnam
7 V. occidentalis ZMMU A5822 Doi Tung Mt., Chiang Rai, MH004406 Poyarkov et al. (2018)
Thailand
8 V. orlovi ZMMU_A5821 Phia Oac-Phia Den NP, Cao MH004404 Poyarkov et al. (2018)
Bang, Vietnam
9 V. orlovi VNMN011309 Phia Oac-Phia Den NP, Cao MW837544 Nguyen et al. 2021
Bang, Vietnam
10 V. vuquangensis VNMN010521 Vu Quang NP, Ha Tinh, MW763028 Hoang et al. (2021)
Vietnam
11 V. vuquangensis VNMN010528 Vu Quang NP, Ha Tinh, MW763029 Hoang et al. (2021)
Vietnam
12 Scaphiophryne boribory FGMV 2000.B24 Unknown MW078574 Scherz et al. (2020)
13 S. boribory Genbank Madagascar: Peyrieras MG020782 Tu et al. (2018)

Phylogenetic analysis

For the phylogenetic analyses, sequences of the five recognized species of Vietnamophryne and the two newly
collected specimens were used in combination with two sequences of Scaphiophryne boribory Vences, Raxworthy,
Nussbaum, & Glaw, 2003 as out-group. Chromas Pro software (Technelysium Pty Ltd., Tewantin, Australia) was
used to edit the sequences, which were aligned using ClustalW (Higgins et al. 1994) implemented in MEGA 7.0
(Kumar et al. 2016) with default settings. We then checked the initial alignments by eye and adjusted them slightly.
Phylogenetic trees were constructed by using maximum likelihood (ML) and Bayesian inference (BI). Prior to
ML and Bayesian analyses, we chose the optimum substitution models for entire sequences using KAKUSAN 4
(Tanabe, 2011) based on the Akaike Information Criterion (AIC). BI tree construction is done via MrBayes version
3.2.7a (Ronquist et al. 2012). The BI summarized two independent runs of four Markov Chains for 10 000 000
generations. A tree was sampled every 100 generations and a consensus topology was calculated for 70,000 trees
after discarding the first 30,001 trees (burnin = 300,010). We checked parameter estimations and convergence using
TRACER version 1.7 (Rambaut et al., 2018). The ML tree was obtained using TreeFinder while the strength of
nodal support in the ML tree was analyzed using non-parametric bootstrapping (MLBS) with 1,000 replicates. We
regarded tree nodes in the ML tree with bootstrap values of 75% or greater as sufficiently resolved (Hillis & Bull,
1993; Huelsenbeck & Hillis, 1993), and nodes with a BPP of 95% or greater as significant in the BI analysis (Leaché
& Reeder, 2002).

Morphological analysis

All measurements were taken from fixed specimens using a digital caliper to the nearest 0.1 mm and subsequently
rounded to a 0.1 mm precision; morphometrics followed Poyarkov et al. (2014, 2018) and Suwannapoom et al.
(2018): (1) snout-vent length (SVL; from tip of snout to cloaca); (2) head length (HL; from tip of snout to hind
border of jaw angle); (3) snout length (SL; from anterior corner of eye to tip of snout); (4) eye length (EL; distance
between anterior and posterior corners of eye); (5) nostril-eye length (N-EL; distance between anterior corner of
eye and nostril center); (6) head width (HW; maximum width of head at level of mouth angles in ventral view);
(7) internarial distance (IND; distance between central points of nostrils); (8) interorbital distance (IOD; shortest
distance between medial edges of eyeballs in dorsal view); (9) upper eyelid width (UEW; maximum distance
between medial edge of eyeball and lateral edge of upper eyelid); (10) forelimb length (FLL; distance between
axilla and elbow) (11) lower arm and hand length (LAL; distance between elbow and tip of third finger); (12) Ua:
Upper arm length (from axilla to tip of finger III); (13) Thigh length (FeL; from vent to knee; (14) tibia length (TL;

A new species of Vietnamophryne from Vietnam Zootaxa 5374 (4) © 2023 Magnolia Press · 507
distance between knee and tibiotarsal articulation); (15) foot and tibiotarsus length (FTL; length from tibiotarsal
joint to end of fourth toe); (16) tympanum length, (TYD; maximum tympanum diameter in horizontal direction);
(17) tympanum-eye distance (TED); (18–21) finger lengths (1–4FLO); (22–26) toe lengths (1–5TOEL).
We obtained comparative morphological data from original descriptions for the following species of
Vietnamophryne reported for Indochina and surrounding parts of Southeast Asia: V. inexpectata (Poyarkov et al.
2018); V. occidentalis (Poyarkov et al. 2018); V. orlovi (Poyarkov et al. 2018); V. vuquangensis (Hoang et al. 2021);
and V. cuongi (Nguyen et al. 2021).

Results

Sequence variation

The 16S rRNA gene fragment consisted of 810 sites: 641 sites were conserved and 168 sites were variable, of which
153 were parsimony-informative. Nucleotide frequencies were A=35.6%, T=23.1%, C=25.1%, and G=16.2%. The
Bayesian and Maximum likelihood analyses produced topologies with –ln L = 2067.99 and 2046.7071, respectively.
The best model selected for BI was the general time reversible model (GTR: Tavaré, 1986) with a gamma shape
parameter (Alpha: 0.529 in BI; Gamma: 0.4039 in ML).

Figure 1. Bayesian inference (BI) tree based on the partial 16S rRNA mitochondrial gene. Values at nodes correspond to
BI/ML support values, respectively.

508 · Zootaxa 5374 (4) © 2023 Magnolia Press NINH ET AL.

 Phylogenetic analyses employing BI method agreed well with those obtained from ML method, thus, only the
Bayesian tree with the most well supported nodes is presented in Figure 1 with support values from both methods.
In the analyses, the newly collected Vietnamophryne specimens were recovered as sister taxon to V. cuongi, V.
occidentalis and V. orlovi with high bootstrap values.
Interspecific uncorrected p-distances of the Vietnamophryne (Table 2) ranged from 2.13% (between V.
orlovi and V. occidentalis) to 6.88% (between V. vuquangensis and V. orlovi). The genetic divergence of the new
Vietnamophryne population from Tuyen Quang Province and its congeners ranged from 2.13% (V. occidentalis) to
6.5% (V. vuquangensis), which was within the genetic distances range of Vietnamophryne species, indicating that
the differentiation between the newly discovered population and other species of Vietnamophryne has reached a
specific level.

TABLE 2. Uncorrected p-distance matrix showing percentage pair-wise genetic divergence of 16S rRNA between
members of the Vietnamophryne species group.
Species - Voucher 1 2 3 4 5 6 7 8 9 10 11 12
number
1. Vietnamophryne
aurantifusca sp. nov.
NH.2023.214
2. Vietnamophryne 0
aurantifusca sp. nov.
NH.2023.215
3. V. cuongi 2.63 2.63
VNMN010358
4. V. cuongi 2.75 2.75 0.13
VNMN010359
5. V. cuongi 2.63 2.63 0 0.13
VNMN010357
6. V. inexpectata 4.5 4.5 5.11 5.26 5.11
MH004403
7. V. occidentalis 2.13 2.13 2.51 2.63 2.51 4.06
MH004406
8. V. orlovi MW837544 2.25 2.25 2.38 2.5 2.38 4.95 2.13
9. V. orlovi MH004404 2.63 2.63 2.75 2.88 2.75 4.8 2.5 0.63
10. V. vuquangensis 6.5 6.5 7.01 7.13 7.01 4.5 6.38 6.5 6.88
MW763029
11. V. vuquangensis 6.5 6.5 7.01 7.13 7.01 4.5 6.38 6.5 6.88 0
MW763028
12. S. boribory 14.18 14.18 14.2 14.3 14.2 14.14 14.32 14.3 14.68 15.91 15.91
MW078574
13. S. boribory 14.3 14.3 14.07 14.18 14.07 13.98 14.2 14.18 14.55 15.79 15.79 0.12
MG020782

Systematics

Vietnamophryne aurantifusca sp. nov.

(Figure 2)

Holotype. IEBR A.5242 (Field number NH.2023.214), adult male, collected in the karst forest of Sinh Long
Commune, Na Hang District, Tuyen Quang Province, Vietnam (22°31’03.4”N; 105°23’18.6”E, at an elevation of
1.056 m a.s.l.) on 25 May 2023 by Nguyen et al.
Paratype. Adult male IEBR A.5243 (Field number NH.2023.215), the same data as the holotype.
Diagnosis. The new species from Tuyen Quang is distinguished from its congeners on the basis of the following

A new species of Vietnamophryne from Vietnam Zootaxa 5374 (4) © 2023 Magnolia Press · 509
characteristics: (1) Body size small SVL (n=2 males, 17.6–18.2 mm); (2) Head wider than long, HW/HL 1.17–1.19;
(3) snout short, round in dorsal view (SL/HW 0.33–0.35), snout length greater than eye length (SL/EL 1.33–1.50);
(4) eye round (EL/HW 0.23–0.25; (5) tympanum small, round, TYD/SVL 0.04–0.05; (6) first finger (F1) well
developed, more than half of second finger (F2) (1FLO/2FLO 0.58–0.64), relative finger lengths: I<II<IV<III,
relative toe lengths: I<II<V<III<IV; (7) tips of all digits round, not expanded in F1–F4, T1, T2, and T5, weakly
expanded in T3 and T4; (8) subarticular tubercles under fingers and toes indistinct; (9) outer metatarsal tubercle
absent, inner metatarsal tubercle small, flattened; (10) dorsal skin relatively smooth with some rounded nodules,
concentrated in the middle of the back, arranged along the length of the back, a prominent ridge along the spine; (11)
orangish-brown with a small triangle shape between eyes; venter orange, with grey marbling, most intense on the
throat, ventral side of limbs dark grey with some whitish marbling (For the measurement see Table 3).

TABLE 3. Measurements (in mm) and proportions of the type series of Vietnamophryne aurantifusca sp. nov.
Field no NH.2023.214/ IEBR A.5242 NH.2024.215/ IEBR A.5243
Holotype Paratype
Sex Male Male
SVL 18.2 17.6
HL 5.8 5.9
SL 2.4 2.3
EL 1.6 1.7
N-EL 1.2 1.1
HW 6.9 6.9
IND 2.0 1.8
IOD 3.0 2.3
UEW 1.1 1.1
FLL 3.6 3.4
LAL 7 7.0
Ua 10.1 10
FeL 8.1 8.2
TL 8.3 8.5
FTL 11.3 11.7
TYD 0.8 0.9
TED 0.8 0.5
1FLO 0.7 0.7
2FLO 1.1 1.2
3FLO 2.1 2.3
4FLO 1.2 1.5
1TOEL 1.7 1.1
2TOEL 3.1 2.6
3TOEL 5.0 5.2
4TOEL 7.5 7.4
5TOEL 4.8 5.0
HW/HL 1.19 1.17
SL/HW 0.35 0.33
SL/EL 1.50 1.35
EL/HW 0.23 0.25
TYD/SVL 0.04 0.05
1FLO/2FLO 0.64 0.58
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 TABLE 3. (Continued)
Field no NH.2023.214/ IEBR A.5242 NH.2024.215/ IEBR A.5243
N-EL/SL 0.50 0.48
TED/SVL 0.04 0.03
LAL/SVL 0.38 0.40
FLL/Ua 0.36 0.34
FeL/TL 0.98 0.97

Figure 2. Dorsal, ventral, and lateral views of the holotype of Vietnamophryne aurantifusca sp. nov (A, B & C). Ventral sides
of the right hand and the left foot with scale bar 2 mm (D & E).

Description of holotype. Habitus stout, body size small (SVL 18.2 mm), head wider than long (HW/HL 1.19),
snout short, obtuse, round in dorsal view, truncate in lateral view (SL/HW 0.35) (Fig. 2), snout length greater than
eye length (SL/EL 1.50); nostril round, lateral, closer to the tip of snout than to eye (N-EL/SL 0.51); dorsal surface
of head slightly convex; canthus rostralis distinct, round; loreal region concave; eyes round medium-sized (EL/HW
0.23); eyes slightly protuberant in dorsal and lateral views; pupil round; tympanum well discernible, circular, small
(TYD/SVL 0.04), located distantly from the eye (TED/SVL 0.04), tympanic rim not elevated above skin of the
temporal area, supratympanic fold present, distinct, glandular (Fig. 2); vomerine teeth and spikes absent; tongue
spatulate and free behind, papillae on tongue absent and vocal sac opening very small and hard to spot, deep in the
palate.
Forelimbs. Short, greater than one-fifth of body length (LAL/SVL 0.38); lower arm shorter than hand, almost
one-third of forelimb length (FLL/Ua 0.36); fingers short, round in cross-section, first finger well developed, more
than half of second finger (1FLO/2FLO 0.64); relative finger lengths: I<II<IV<III (Fig. 2D); webbing absent; dermal
fringes present, weakly developed. Tip of the first finger rounded, slightly tapered. Tips of fingers II–V rounded,
not dilated, finger disks absent, terminal grooves absent; longitudinal furrow on dorsal surface of fingers absent;
subarticular tubercles under fingers indistinct; nuptial pad absent; two palmar tubercles: inner palmar tubercle small,
rounded; outer palmar tubercle rounded; palmar surface smooth, supernumerary palmar tubercles absent.
Hindlimbs. Short and thick, thigh length less than half of the snout-vent length (FeL/SVL 0.45); tibiotarsal

A new species of Vietnamophryne from Vietnam Zootaxa 5374 (4) © 2023 Magnolia Press · 511
articulation of adpressed limb reaching eye level; femur length nearly equal to tibia length (FeL/TL 0.97); relative
toe lengths: I<II<V<III<IV; tarsus smooth, tarsal fold absent; tips of toes rounded, first toe slightly tapered, tips of
toes III and IV slightly dilated (Figure. 2E), terminal grooves absent; toes rounded in cross-section; webbing absent
between all toes; dermal fringes on toes present, weakly developed; subarticular tubercles under toes ovoid, slightly
flattened; single metatarsal tubercle: inner metatarsal tubercle rounded, flattened.
Skin texture. Dorsal skin relatively smooth with some rounded nodules, concentrated in the middle of the back,
arranged along the length of the back, a prominent ridge along the spine, dorsal surfaces of the forearm, posterior
and lateral parts of the dorsum, sacral area, and dorsal surfaces of hindlimbs shagreened; upper eyelids slightly
harsh, supratympanic folds with low glandular ridges; ventral sides of the trunk, head and limbs completely smooth;
well developed distinct dermal ridge present on midline of head and dorsum, running from between eyes to vent.
Color of holotype in life. Dorsum orangish-brown, posteriorly darker, tubercles dark brown, densely scattered
on dorsal surfaces of head, body, and limbs (Fig. 2A, C); posterior parts of dorsum of hindlimbs dark brown with
black pustules; canthus rostralis ventrally dark brown; loreal region dark with some whitish mottling on lower jaw
and mouth; upper eyelids dark brown; dorsal surfaces of forearms orange brown interspersed with greyish brown;
supratympanic fold dark-brown with some whitish pustules; ventrally orange, fainter on chest, throat, and ventral
surfaces of limbs, and with grey marbling, concentrated on the throat and ventral surfaces of limbs; ventral sides of
fingers and toes dark grey with some whitish marbling. Pupil rounded, black, iris uniform dark brown.
Color of holotype in preservative: Dorsal surface changed from brown to grey scattered with dark-brown
tubercles and pustules; ventral surface of body faded completely to creamy white with dark-grey marbling denser
on the throat, hands, tibia and feet.

Figure 3. Dorsal view of Vietnamophryne aurantifusca sp. nov., holotype (NH.2023.214) A and B; paratype (NH.2023.215)
C and D after fixation.

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 TABLE 4. Selected diagnostic characteristics for the species in the genus Vietnamophryne. ND = No data.
Ratio head Ratio snout
Dorsolateral Tympanum Ventral
Species Sex SVL width and length and eye Finger 1 length Dorsal skin texture
white spots length coloration
head length length
Ventrally
orange
Snout length Well developed,
Head wider Small tympanum coloration Dorsal skin relatively
Vietnamophryne greater than eye more half of second
Male 17.55–18.24 Present than long (HW/ (TYD/SVL 0.04– with grey smooth, with some
aurantifusca sp. length (SL/EL finger (1FLO/
HL 1.17–1.19) 0.05) marbling rounded nodules
nov. 1.33–1.45) 2FLO 0.59–0.66)
concentrate on
the throat
Female ND ND ND ND ND ND ND ND
Male ND ND ND ND ND ND ND ND

A new species of Vietnamophryne from Vietnam

Well developed, Ventrally
Vietnamophryne Snout length Mostly smooth skin
Head wider Large tympanum about half of milky-white
cuongi greater than eye on dorsum with
Female 16.9–18.1 Present than long (HW/ (TYD/SVL 0.08– second finger coloration
length (SL/EL somewhat enlarged
HL 1.10–1.23) 0.09) (1FLO/2FLO with gray
1.33–1.42) flat pustules
0.43–0.62) marbling
Smooth anteriorly,
Ventrally
Snout length Well developed, somewhat shagreened
Head longer Smaller bright lemon-
greater than eye half of second posteriorly with small
Male 15.4 Present than wide (HW/ tympanum (TYD/ yellow with
length (SL/EL finger (1FL/2FL flat pustules loosely
HL 0.87) SVL 0.06) dark brown
1.39) 0.50) scattered on posterior
marbling
dorsum
V. orlovi
Smooth anteriorly,
Ventrally
Snout length Well developed, somewhat shagreened
Head longer Smaller bright lemon-
greater than eye half of second posteriorly with small
Female 17.1 Present than wide (HW/ tympanum (TYD/ yellow with
length (SL/EL finger (1FLO/ flat pustules loosely
HL 0.98) SVL 0.07). dark brown
1.31) 2FLO 0.50) scattered on posterior
marbling
dorsum
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513
 TABLE 4. (Continued)
Ratio head Ratio snout
Dorsolateral Tympanum Ventral
Species Sex SVL width and length and eye Finger 1 length Dorsal skin texture
white spots length coloration
head length length

Ventrally
Tympanum Well developed,
Head wider Snout length ivory-lemon to Shagreened with
relatively faint, less than half of
than long, greater than eye lemon-yellow small flat pustules
Male 14.1–14.9 Present comparatively second finger
(HW/HL 1.17– length (SL/EL with weak loosely scattered on
small, rounded, (1FLO/2FLO
1.24) 1.19–1.28) dark-brown dorsum
0.05–0.06 of SVL 0.43–0.49)
marbling
V. vuquangensis

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Well developed, Ventrally
Head wider Snout length Tympanum Mostly smooth,
less than half of bright lemon-
than long, greater than eye comparatively slightly shagreened
Female 15.2–17.3 Present second finger yellow with
(HW/HL 1.10– length (SL/EL small, rounded, posteriorly, lacking
(1FLO/2FLO weak dark-
1.31) 1.03–1.30) 0.05–0.06 of SVL enlarge tubercles
0.44–0.50) brow marbling

Ventrally Warty skin on

Snout length Rudimentary,
Head as long as grayish-beige posterior and
comparable eye Larger tympanum slightly protuberant
Male 14.2 Present wide, (HW/HL coloration shagreened skin
V. inexpectata length (SL/EL (TYD/SVL 0.09) as nub (1FLO/
1.01) with gray on anterior parts of
0.95) 2FLO 0.30)
marbling dorsum
Female ND ND ND ND ND ND ND ND
Ventrally
Completely smooth,
Head less Snout length Tympanum bright
Well developed, posteriorly with
longer than shorter than eye comparatively orange-red
Male 20.5 Absent half of F2 length loosely scattered
V. occidentalis wide (HW/HL length (SL/EL small, round, 0.05 with sparse
(1FL/2FL 0.43) small flat tubercles
0.99) 0.84) of SVL dark brown
present
marbling
Female ND ND ND ND ND ND ND ND

NINH ET AL.
 Variation. The holotype IEBR A.5242 has more tubercles on dorsum than the paratype IEBR A.5243.
Etymology. Specific epithet is based on the Latin word for orange (aurantium) and brown (fuscus) and refers to
the orange brown colouration of the new species. As for the common English name of the new species, we propose
Orange-brown Dwarf Frog and the common name in Vietnamese as “Nhái lùn nâu cam”.
Distribution. Vietnamophryne aurantifusca sp. nov. is currently known only from the type locality in Tuyen
Quang Province, Vietnam.
Comparisons. Comparative morphological characteristics are given in Table 4. Vietnamophryne aurantifusca sp.
nov. can be distinguished from the other Vietnamophryne species by a combination of the following characteristics:
from V. cuongi by having dorsal skin relatively smooth, with some rounded nodules, concentrated in the middle of
the back vs. mostly smooth skin on dorsum with somewhat enlarged flat pustules in V. cuongi; Dorsum orangish-
brown with a grey patch in triangle shape between eyes; ventral side orange, with grey marbling in Vietnamophryne
aurantifusca sp. nov. vs. dorsally black brown, pustules on lateral dorsum white-brown and ventral side milky-
white with grey marbling in V. cuongi (Nguyen et al. 2021).
From V. inexpectata by having a larger size in males (SVL 17.6–18.2 mm vs. 14.2 mm in V. inexpectata); head
wider than long vs head as long as wide (HW/HL 1.17–1.19 vs. 1.01 in V. inexpectata); snout length greater than
eye length (SL/EL 1.35–1.50 vs. 0.95 in V. inexpectata); a smaller tympanum (TYD/SVL 0.04–0.05 vs. 0.09 in V.
inexpectata); first finger more developed (1FLO/2FLO 0.58–0.64 vs. 0.30 in V. inexpectata); ventral surface orange
(vs. greyish-beige coloration in V. inexpectata; dorsum relatively smooth with some rounded nodules, arranged
along the length of the back, a prominent ridge along the spine (vs. warty skin on posterior and shagreened skin on
anterior parts of dorsum in V. inexpectata) (Poyarkov et al. 2018).
From V. occidentalis by having a smaller size (SVL 17.6–18.2 mm vs. 20.5 in V. occidentalis); white spots on
dorsolateral region (vs. absent in V. occidentalis); snout length greater than eye length (SL/EL 1.33–1.42 vs. 0.84 in
V. occidentalis; first finger well developed (1FL/2FL 0.59–0.66 vs 0.43 in V. occidentalis); ventral surface orange
with grey marbling (vs. bright orange-red with sparse dark brown marbling in V. occidentalis) (Poyarkov et al.
2018).
From V. orlovi by having a greater size in males (SVL SVL 17.6–18.2 mm vs. 15.4 mm in V. orlovi); head
wider than long vs head longer than wide (HW/HL 1.17–1.19 vs. 0.87 in V. orlovi); a smaller tympanum (TYD/SVL
0.04–0.05 vs. 0.06 in V. orlovi); first finger more developed (1FLO/2FLO 0.58–0.64 vs. 0.50 in V. orlovi); ventral
surface orange with grey marbling (vs. bright lemon-yellow with dark brown marbling in V. orlovi) (Poyarkov et
al. 2018).
From V. vuquangensis by having a greater size (SVL 17.6–18.2 mm vs.14.1–14.9 mm in V. vuquangensis); first
finger more developed (1FLO/2FLO 0.58–0.64 vs. 0.43–0.49 in vs. V. vuquangensis); ventral surface orange with
grey marbling (vs. ivory-lemon to lemon-yellow with weak dark-brown marbling in V. vuquangensis). Dorsum
orangish-brown, posteriorly darker, tubercles dark brown densely scattered in Vietnamophryne aurantifusca sp.
nov. (vs. dorsum reddish-grey on black tone, black markings mixed with small, densely scattered speckles in V.
vuquangensis) (Hoang et al. 2021).

Natural history

Specimens of the new species were found at night on the ground, under decaying leaves on the edge of the trails in
regenerating secondary forests at elevations of about 1.000 m a.s.l. (Fig. 4), covered with rotten leaves under the
canopy of trees belonging to the Polypodiaceae family and Commelina genus. The air temperature at the time of the
finding was not recorded but was 30oC at 20:00. Other species of amphibians that were found at night in the same
habitat were Gracixalus gracilipes (Bourret, 1937), Kurixalus sp. and Polypedates sp.

A new species of Vietnamophryne from Vietnam Zootaxa 5374 (4) © 2023 Magnolia Press · 515
Figure 4. Habitat at the type locality of Vietnamophryne aurantifusca sp. nov.

Discussion

Vietnam is famous for its high level of species richness of amphibians with more than 250 recorded species (Frost
2023). Most of the new discoveries are because of the splitting of complex taxa due to integrative taxonomic analyses.
Some of the new species are of small size and previously escaped the attention of researchers. Vietnamophryne is
such a newly discovered amphibian genus (Poyarkov et al. 2018), of which so far six species have been recorded,
including the new species Vietnamophryne aurantifusca. Five species are known only from Vietnam, making this
country a hot spot for dwarf frog diversity. Until now each of these species was only recorded from one study site
(Fig. 5).
In addition, the type locality of V. aurantifusca is about 50 km far from Phia Oac-Phia Den NP (type locality
of V. orlovi), 145 km far from Ba Vi NP (type locality of V. cuongi), 450km from Vu Quang NP (type locality of
V. vuquangenis), 650 km from Chiang Rai, Thailand (type locality of V. occidentalis), and the furthest is Kon Chu
Rang Nature Reserve about 950 km (type locality of V. inexpectata). Because the locality of V. aurantifusca is
relatively close to that of V. orlovi and the genus is distributed across Vietnam as well as Thailand, there may be a
number of undiscovered species located close to each other in the region. Therefore, it is possible that the number of
species in this genus is underestimated, and future field works in the region may uncover new species.
Previous studies showed that their distribution is restricted to altitudes ranging from 600 to 1200 m a.s.l., where
they can be found in moist soil with decaying vegetation and rotting roots. So far, all discoveries in Vietnam took
place from May to June, which is the beginning of the rainy season. Vietnamophryne is clearly adapted to living
on relatively moist soil with conspicuous leaf litter. Certainly, more research is needed to better understand the
diversity and biology of Vietnamophryne species.

516 · Zootaxa 5374 (4) © 2023 Magnolia Press NINH ET AL.

Figure 5. Type locality (red circle) of Vietnamophryne aurantifusca sp. nov. in Sinh Long Commune, Na Hang District,
Tuyen Quang Province, Northeastern Vietnam and distribution of Vietnamophryne species.

Acknowledgements

We are grateful to the directorates of the Forest Protection Department of Tuyen Quang Province and Na Hang
District for supporting our fieldwork and issuing relevant permits. We thank Nguyen T.N. (IGR) for her assistance
in the laboratory work. This research is funded by the Vietnam Academy of Science and Technology (VAST) under
Grant Number THTETN.07/23-24.

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