DSpace at VNU: First Record of the Genus Oreolalax (Anura: Megophryidae) from Vietnam with Description of a New Species

First Record of the Genus Oreolalax (Anura: Megophryidae) from Vietnam with
Description of a New Species
Author(s): Truong Q. Nguyen , Trung M. Phung , Minh D. Le , Thomas Ziegler , and Wolfgang Böhme
Source: Copeia, 2013(2):213-222. 2013.
Published By: The American Society of Ichthyologists and Herpetologists
DOI: />URL: />
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Copeia 2013, No. 2, 213–222

First Record of the Genus Oreolalax (Anura: Megophryidae) from Vietnam
with Description of a New Species
Truong Q. Nguyen1, Trung M. Phung2, Minh D. Le3,4,5, Thomas Ziegler6, and
Wolfgang Bo
¨ hme7
The genus Oreolalax is reported from Vietnam for the first time and a new species is described based on morphological
differences, molecular divergence, and phylogenetic placement. Morphologically, the new species is distinguishable
from its congeners on the basis of a combination of the following diagnostic characters: size small; tympanum hidden;
toes with webbing at base; dorsum with distinct, round, spiny warts; flanks with white, spiny spots; belly and lower
surface of limbs smooth, with dark marbling; interorbital region without dark triangular pattern; upper surface of
thigh with dark bars; male with black spines present on margin of lower lip, spinal patches on chest small with fine

spines, nuptial spines on fingers small, and without vocal sacs. In phylogenetic analyses, the new species is
unambiguously nested within the genus Oreolalax.

T

HE genus Oreolalax was created by Myers and Leviton
(1962) for four species of Scutiger described by Liu
(1943, 1947) from western China. Dubois (1980,
1987) treated Oreolalax as a subgenus of Scutiger. Ohler and
Dubois (1992) also assigned Megalophrys weigoldi Vogt, 1924
to the genus Scutiger (subgenus Aelurolalax). However, the
placement of this species in the genus Oreolalax was
mentioned by Fei (1999) and subsequently followed by
Delorme et al. (2006), Wei et al. (2007, 2009), and Fei et al.
(2009, 2010). On the other hand, Oreolalax was recently
considered as a distinct genus based on both morphological
and phylogenetic data (e.g., Wu et al., 1993; Zhao and Adler,
1993; Fu and Murphy, 1997; Zheng et al., 2004; Delorme et
al., 2006; Fu et al., 2007; Wei et al., 2007). Members of the
genus Oreolalax are characterized by having maxillary teeth
short; pupils vertically elliptical; tongue deeply notched
posteriorly; eustachian tubes with large, rounded openings;
tympana hidden beneath skin; fingers free of webbing; toes
with some webbing; outer metatarsals united; dorsum
glandular with numerous warts; males with large, flat, but
prominent pectoral and axillary glands; and palmar tubercles large, flat, and not extending onto first metatarsal
(Myers and Leviton, 1962).
The genus Oreolalax currently consists of 17 species and so
far is known only from southwestern China (Frost, 2011).
Frost (2011) assumed that it might occur in Laos and

Vietnam; however, no voucher specimen has been reported
previously. During recent field work in the Hoang Lien
Mountain Range, we discovered a species of frog which
resembled Oreolalax, but it was not assignable to any of the
named species so far. Herein, we describe this species as new,
assign it to Oreolalax based on morphological characters and
phylogenetic analysis of nucleotide sequence data, and
report the genus from Vietnam for the first time.
1

MATERIALS AND METHODS
Sampling.—A field survey was conducted in April 2010 in the
Hoang Lien Mountain Range, Sa Pa District, Lao Cai
Province, northern Vietnam. Specimens were collected by
T. M. Phung. Samples of muscle tissue were preserved
separately in 95% ethanol and voucher specimens were
fixed in approximately 80% ethanol, and then later
transferred to 70% ethanol for permanent storage. DNA
sequence data from our new samples were combined with a
previously collected mitochondrial sequence dataset for
species of Oreolalax and relevant out-groups, including
Leptobrachium boringiae, L. xanthospilum, Megophrys omeimontis, and Scutiger chintingensis (Zheng et al., 2004; Fu et al.,
2007; Table 1). Sequences of Oreolalax granulosus, O. puxiongensis, O. weigoldi, and O. xiangchengensis are unobtainable from GenBank; therefore, these species were excluded
from our phylogenetic analysis in this study. Specimens
referred to in this paper are deposited in the collections of
the Chengdu Institute of Biology (CIB), Chinese Academy of
Sciences, Sichuan, China; the Institute of Ecology and
Biological Resources (IEBR), Hanoi, Vietnam; the Institute
of Zoology (IOZ), Chinese Academy of Sciences, Beijing,
China; the Royal Ontario Museum (ROM), Toronto,

Canada; the Kunming Institute of Zoology (KIZ), Chinese
Academy of Sciences, Yunnan, China; and the Zoologisches Forschungsmuseum Alexander Koenig (ZFMK),
Bonn, Germany.
Nucleotide sequencing and phylogenetic analyses.—We used Le
et al.’s (2006) protocols for extraction, amplification, and
DNA sequencing. Two fragments of the mitochondrial genes
12S and 16S were amplified using two pairs of primers,
L1991 + H1478 for 12S (Kocher et al., 1989) and 16Sar +
16Sbr for 16S (Palumbi et al., 1991). After sequences were

Department of Vertebrate Zoology, Institute of Ecology and Biological Resources, Vietnam Academy of Science and Technology, 18 Hoang
Quoc Viet Road, Hanoi, Vietnam; Present address: Department of Terrestrial Ecology, Cologne Biocenter, University of Cologne, Zu
¨ lpicher
Strasse 47b, D-50674 Cologne, Germany; E-mail: Send reprint requests to this address.
2
Dong Khoi 9A, Tam Hiep, Bien Hoa, Dong Nai Province, Vietnam; E-mail:
3
Centre for Natural Resources and Environmental Studies, Hanoi National University, 19 Le Thanh Tong, Hanoi, Vietnam; E-mail:

4
Faculty of Environmental Sciences, Hanoi University of Science, Vietnam National University, 334 Nguyen Trai Road, Hanoi, Vietnam.
5
Department of Herpetology, American Museum of Natural History, Central Park West at 79th Street, New York 10024-5192.
6
AG Zoologischer Garten Ko
¨ ln, Riehler Strasse 173, D-50735 Cologne, Germany; E-mail:
7
Herpetology Section, Zoologisches Forschungsmuseum Alexander Koenig, Adenauerallee 160, D-53113, Bonn, Germany; E-mail:

Submitted: 15 February 2012. Accepted: 15 November 2012. Associate Editor: B. Stuart.

DOI: 10.1643/CH-12-021
F 2013 by the American Society of Ichthyologists and Herpetologists


214

Copeia 2013, No. 2

Table 1. Samples Used in Molecular Analyses.

Species information
Oreolalax chuanbeiensis
Oreolalax jingdongensis
Oreolalax liangbeiensis
Oreolalax lichuanensis
Oreolalax major
Oreolalax multipunctatus
Oreolalax nanjiangensis
Oreolalax omeimontis
Oreolalax pingii
Oreolalax popei
Oreolalax rhodostigmatus
Oreolalax rugosus
Oreolalax schmidti
Oreolalax sp. (China)
Oreolalax sterlingae, new
species (Vietnam)
Oreolalax sterlingae, new
species (Vietnam)
Oreolalax xiangchengensis

Leptobrachium boringiae
Leptobrachium xanthospilum
Megophrys omeimontis
Scutiger chintingensis

GenBank no. (12S)

GenBank no. (16S)

Locality

Voucher

EF397266
EF544236
EF397253
EF544237
EF397252
EF397268
EF397265
EF397264
EF397259
—
EF397248
EF397254
EF397258
EF397256
KC569979

EF397266

EF544236
EF397253
EF544237
EF397252
EF397268
EF397265
EF397264
EF397259
EF397267
EF397248
EF397254
EF397258
EF397256
KC569981

Mao Xian, Sichuan, China
—
Puxiong, Sichuan, China
—
Hongya, Sichuan, China
Omei, Sichuan, China
Nan Jiang, Sichuan, China
Omei, Sichuan, China
Xi Chang, Sichuan, China
Penxian, Sichuan, China
Da Fang, Guizhou, China
Shi Mian, Sichuan, China
Da Yi, Sichuan, China
Pengxian, Sichuan, China
Sa Pa, Lao Cai, Vietnam

CIB-XM 074
IOZCAS 2691–2692
IOZCAS 3796
IOZCAS 2675
ROM 40452
ROM 40463
CIB-XM 804
CIB-XM 439
CIB-XM 980
CIB-XM 107
CIB-ZYCA 746
CIB-XM 340
CIB-XM 417
CIB-XM 092
IEBR A.2012.1

KC569980

KC569982

Sa Pa, Lao Cai, Vietnam

ZFMK 92830

EF397250
EF397247
EF397245
EF397242
EF397269

EF397250
EF397247
EF397245
EF397242
AY526209

Li Jiang, Yunnan, China
Leishan, Guizhou, China
Gia Lai, Vietnam
Omei, Sichuan, China
Hongya, Sichuan, China

CIB-3LW 008
CIB-XM 594
ROM 32184
ROM 40462
ROM 39065 (12S)
/XM 1045 (16S)

aligned by Clustal X v2 (Thompson et al., 1997), data were
analyzed using maximum parsimony (MP) and Bayesian
analysis (BA), as implemented in PAUP*4.0b10 (Swofford,
2001) and MrBayes v3.2 (Huelsenbeck and Ronquist, 2001),
respectively. Settings for these analyses followed Le et al.
(2006), except that the number of generations in the
Bayesian analysis was increased to 13107. The optimal
model for nucleotide evolution was set to GTR+I+C as
selected by Modeltest v3.7 (Posada and Crandall, 1998).
Nodal support was evaluated using Bootstrap replication

(BP) as calculated in PAUP and posterior probability (PP) in
MrBayes v3.2. Uncorrected pairwise divergences were calculated in PAUP*4.0b10.
Morphological analysis.—We compared the new species with
other members of the genus (see Material Examined) and
data obtained from the literature (Liu and Hu, 1960; Liu et
al., 1979; Tian, 1983; Fei and Huang, 1983; Yang et al., 1983;
Fei et al., 1992, 1999, 2009, 2010; Ohler and Dubois, 1992;
Fu and Murphy, 1997; Wei et al., 2009). Measurements were
taken with digital callipers to the nearest 0.1 mm. The
following abbreviations were used: SVL: snout–vent length,
HL: head length (from posterior margin of lower jaw to the
tip of snout), HW: maximum head width (across angle of
jaws), SNL: snout length (from anterior corner of eye to the
tip of snout), NS: distance from nostril to the tip of snout,
EN: distance from anterior corner of eye to the nostril, IN:
internarial distance, IOD: interorbital distance, ED: eye
diameter, UEW: maximum width of upper eyelid, DAE:
distance between anterior corners of eyes, DPE: distance
between posterior corners of eyes, MAE: distance between
posterior margin of lower jaw and anterior corner of eye,
MPE: distance between posterior margin of lower jaw and
posterior corner of eye, MN: distance from posterior margin

of lower jaw to the nostril, FLL: forelimb length (from axilla
to elbow), HAL: hand length (from elbow to the tip of third
finger), FFL: first finger length, TFL: third finger length, IPT:
inner palmar tubercle length, OPT: outer palmar tubercle
length, FeL: femur length (from vent to knee), TbL: tibia
length (from knee to tarsus), FoL: foot length (from tarsus to
the tip of fourth toe), FTL: fourth toe length, IMT: inner

metatarsal tubercle length, OMT: outer metatarsal tubercle
length, ParL: parotoid length (from posterior corner of eye to
the end of parotoid gland), SPW: maximum width of spinal
patch on chest, CW: chest width (distance between axillae).
We followed Wei et al. (2009) to define the size of spinal
patches on chest: small if the ratio of SPW/CW ,0.3 and large
if this ratio .0.4. Photos of the preserved specimen were
taken with a digital microscope Keyence VHX-500F.
RESULTS
Phylogenetic analyses.—The combined matrix contains 867
aligned characters, 375 of 12S and 492 of 16S. MP analysis of
the dataset recovered nine most parsimonious trees with 609
steps (CI 5 0.69, RI 5 0.63). One of the nine trees is shown
in Figure 1. Fifty percent of the in-group nodes in our
preferred tree received strong support (Bootstrap value $
70%; Hillis and Bull, 1993), and the species from Sa Pa is
unambigously nested in the genus Oreolalax (Bootstrap
value 5 97). In the Bayesian analysis, –lnL scores reached
stationarity after 11,000 generations in both runs. The
Bayesian topology is generally in agreement with the MP
topology, but we note a few significant discrepancies. In the
Bayesian topology, the species of Oreolalax from Sa Pa along
with other clades, including O. multipunctatus and other
species, O. jingdongensis and others, and O. lichuanensis and
others, are unresolved. In addition, the positions of O.


Nguyen et al.—New Oreolalax from Vietnam

215

Fig. 1. The maximum parsimony tree based on partial 12S and 16S genes. This is one of nine most parsimonious trees (TL 5 609, CI 5 0.69, RI 5
0.63). The dataset includes 867 aligned characters of which 156 are potentially parsimony informative. Numbers above and below branches are
bootstrap (.50%) values and Bayesian posterior probabilities (.95%), respectively.

chuanbeiensis and O. nanjiangensis are interchanged, and O.
liangbeiensis and O. major become sister with O. rugosus
replacing the position of O. liangbeiensis as shown in
Figure 1. Both MP and BA analyses strongly support the
monophyly of the genus Oreolalax including the new species
(BP 5 97, PP 5 98), and demonstrate that the Vietnamese
species is not closely related to any other members of the

genus (Fig. 1). This species is largely divergent from others
within Oreolalax in terms of genetic distance, with the
minimum pairwise divergence of approximately 5% in 16S,
the fastest evolving marker in this study (Table 2).
Morphological comparisons.—The new Vietnamese species of
Oreolalax differs from previously named species as follows:

Table 2. Uncorrected (‘‘p’’) Distance Matrix Showing Pairwise Genetic Divergence between Vietnamese Oreolalax and Other Species in the Genus
(except O. granulosus, O. puxiongensis, O. weigoldi, and O. xiangchengensis).

Oreolalax sterlingae, new
species
Species name
Oreolalax
Oreolalax
Oreolalax
Oreolalax

Oreolalax
Oreolalax
Oreolalax
Oreolalax
Oreolalax
Oreolalax
Oreolalax
Oreolalax
Oreolalax
Oreolalax
Oreolalax

chuanbeiensis
jingdongensis
liangbeiensis
lichuanensis
major
multipunctatus
nanjiangensis
omeimontis
pingii
popei
rhodostigmatus
rugosus
schmidti
sp. (China)
xiangchengensis

(16S)
0.073

0.058
0.054
0.073
0.054
0.068
0.068
0.066
0.064
0.073
0.073
0.048
0.066
0.056
0.056

Oreolalax sterlingae, new
species
(12S)
0.046
0.022
0.022
0.035
0.022
0.051
0.046
0.040
0.022
—
0.051
0.022

0.024
0.022
0.022

Oreolalax sterlingae, new
species
(total)
0.061
0.042
0.040
0.056
0.040
0.061
0.059
0.055
0.046
0.072
0.063
0.036
0.048
0.041
0.041


216

from O. chuanbeiensis by having a smaller size (SVL 36.8 mm
[n 5 1] vs. 47.5–56.3 mm [n 5 20] in males and 44.6 mm [n
5 1] vs. 55.5–59.0 mm [n 5 3] in females), white spots on
flanks present (vs. absent in O. chuanbeiensis), and fringes

along the side of toes absent (vs. present in O. chuanbeiensis);
from O. granulosus by having a smaller size (SVL 36.8 mm [n
5 1] vs. 48.6–61.5 mm [n 5 16] in males and 44.6 mm [n 5
1] vs. 57.4–60.0 mm [n 5 3] in females), spinal patches on
chest smaller (vs. very large in O. granulosus), toes with
webbing at base only (vs. webbing well developed in O.
granulosus), and white spots on flanks present (vs. absent in
O. granulosus); from O. jingdongensis by having a smaller size
(SVL 36.8 mm [n 5 1] vs. 48.1–60.3 mm [n 5 16] in males),
spines on chest small (vs. large in O. jingdongensis), nuptial
spines on fingers small (vs. large in O. jingdongensis),
webbing on toes basal (vs. 1/3 webbed in O. jingdongensis),
and triangular pattern between the eyes absent (vs. present
in O. jingdongensis); from O. liangbeiensis by having a smaller
size (SVL 36.8 mm [n 5 1] vs. 45.1–56.3 mm [n 5 20] in
males and 44.6 mm [n 5 1] vs. 56.0–65.7 mm [n 5 8] in
females), spinal patches on chest small (vs. very large in O.
liangbeiensis), and belly with dark marbling (vs. immaculate
in O. liangbeiensis); from O. lichuanensis by having a smaller
size (SVL 36.8 mm [n 5 1] vs. 50.0–64.8 mm [n 5 2] in males
and 44.6 mm [n 5 20] vs. 57.3–62.2 mm [n 5 4] in females),
spines on chest small (vs. large in O. lichuanensis), nuptial
spines on fingers small (vs. large in O. lichuanensis), and
white spots on flanks present (vs. absent in O. lichuanensis);
from O. major by having a smaller size (SVL 36.8 mm [n 5 1]
vs. 59.2–68.7 mm [n 5 6] in males and 44.6 mm [n 5 1] vs.
65.0–70.0 mm [n 5 2] in females), spinal patches on chest
small (vs. very large in O. major), webbing on toes less
developed (at base vs. 1/3–2/3 webbed), fringes along the
side of toes absent (vs. present in O. major), and white spots

on flanks present (vs. absent in O. major); from O. multipunctatus by having a smaller size (SVL 36.8 mm [n 5 1] vs.
47.4–49.8 mm [n 5 4] in males), spines on chest small (vs.
large in O. multipunctatus), nuptial spines on fingers small
(vs. large in O. multipunctatus), white spots on flanks present
(vs. absent in O. multipunctatus), and round black spots on
dorsum absent (vs. present in O. multipunctatus); from O.
nanjiangensis by having a smaller size (SVL 36.8 mm [n 5 1]
vs. 52.6–60.0 mm [n 5 10] in males and 44.6 mm [n 5 1] vs.
53.3–58.2 mm [n 5 8] in females), spines on margin of lower
jaw present (vs. absent in O. nanjiangensis), spines on chest
small (vs. large in O. nanjiangensis), nuptial spines on fingers
small (vs. large in O. nanjiangensis), belly with dark marbling
(vs. immaculate in O. nanjiangensis), and white spots on
flanks present (vs. absent in O. nanjiangensis); from O.
omeimontis by having a smaller size (SVL 36.8 mm [n 5 1] vs.
49.5–58.4 mm [n 5 15] in males and 44.6 mm [n 5 1] vs.
51.2–56.1 mm [n 5 3] in females), spines on margin of lower
jaw present (vs. absent in O. omeimontis), vocal sac absent in
males (vs. present in O. omeimontis), triangular pattern
between eyes absent (vs. present in O. omeimontis), and
white spots on flanks present (vs. absent in O. omeimontis);
from O. pingii by having a dorsum with large round warts
(vs. small warts in O. pingii), belly with dark marbling (vs.
immaculate in O. pingii), and dark bars on upper side of
limbs present (vs. absent in O. pingii); from O. popei by
having a smaller size (SVL 36.8 mm [n 5 1] vs. 60.0–69.0 mm
[n 5 20] in males and 44.6 mm [n 5 1] vs. 51.5–67.0 mm [n
5 10] in females), spines on chest small (vs. large in O.
popei), nuptial spines on fingers small (vs. large in O. popei),

Copeia 2013, No. 2

dark spots with light centers on dorsum absent (vs. present
in O. popei), and white spots on flanks present (vs. absent in
O. popei); from O. puxiongensis by having a dorsum with
round warts (vs. back with strongly developed spiny ridges
in O. puxiongensis), belly with marbling (vs. immaculate in
O. puxiongensis), nuptial spines on fingers small (vs. large in
O. puxiongensis), and triangular pattern between eyes absent
(vs. present in O. puxiongensis); from O. rhodostigmatus by
having a smaller size (SVL 36.8 mm [n 5 1] vs. 57.5–73.5 mm
[n 5 7] in males and 44.6 mm [n 5 1] vs. 62.4–70.6 mm [n 5
2] in females), tympanum hidden (vs. distinct in O.
rhodostigmatus), and large orange round warts on axilla
and at the back of thigh absent (vs. present in O.
rhodostigmatus); from O. rugosus by having toes with less
developed webbing (at base vs. 1/4 webbed in O. rugosus),
toes with lateral fringes absent (vs. present in O. rugosus),
and light spots on flanks present (vs. absent in O. rugosus);
from O. schmidti by having a belly with marbling (vs.
immaculate in O. schmidti), nuptial spines on fingers small
(vs. large in O. schmidti), and dark triangular pattern
between the eyes absent (vs. present in O. schmidti); from
O. weigoldi by having a smaller size (SVL 36.8 mm [n 5 1] vs.
58.2 mm [n 5 1] in males), two clearly separated spinal
patches on chest (vs. one large spinalpatch in O. weigoldi),
toes with webbing at base (vs. well developed in O. weigoldi),
and belly smooth (vs. granular in O. weigoldi); from O.
xiangchengensis by having a smaller size (SVL 36.8 mm [n 5
1] vs. 45.4–50.6 mm [n 5 10] in males and 44.6 mm [n 5 1]

vs. 54.3–61.4 mm [n 5 10] in females), spinal patches on
chest small (vs. very large in O. xiangchengensis), and toes
with webbing at base (vs. almost completely webbed in O.
xiangchengensis; Table 3).
Based on the results of the molecular analyses and
morphological comparisons, we argue that the Vietnamese
specimens represent a new species, which is assigned to the
genus Oreolalax and is described below.
Oreolalax sterlingae, new species
Holotype.—IEBR A.2012.1, adult male, northern Vietnam,
Lao Cai Province, Sa Pa District, Hoang Lien Mountain,
2900 m, Phung My Trung, 30 April 2010.
Paratype.—ZFMK 92830, adult female, same collection data
as the holotype.
Diagnosis.—The new species differs from its congeners by a
combination of the following morphological characters: size
small (SVL 36.8 mm in male, 44.6 mm in female);
tympanum hidden; toes with webbing at base; dorsum with
distinct round spiny warts; flanks with white spiny spots;
belly and lower surface of limbs smooth, with dark
marbling; interorbital region without dark triangular pattern; upper surface of thigh with three wide dark bars; male
with black spines present on margin of lower lip, spinal
patches on chest small with fine spines, nuptial spines on
fingers small, and without vocal sacs.
Description of holotype.—Size small (SVL 36.8 mm). Head
wider than long (HL 13.3 mm, HW 13.7 mm); snout
protruding, rounded anteriorly, snout length (SNL 6.1 mm)
greater than horizontal diameter of eye (ED 5.1 mm);
interorbital region flat, wider (IOD 4.3 mm) than upper
eyelid (UEW 3.3 mm); canthus rostralis distinct, loreal

region concave; nostril oval, on lateral side, lying midway


SVL (mm)
Males
Females
Tympanum (hidden 5 0, distinct 5 1)
Wart (round 5 0, elongated ridge 5 1)
Belly (smooth 5 0, granular 5 1)
Belly (immaculate 5 0, with marbling 5 1)
Triangular pattern between eyes
(absent 5 0, present 5 1)
Toe webbing
Vocal sac (absent 5 0, present 5 1)
Spines on lip margin (absent 5 0, present 5 1)
Spinal patches on chest (small 5 0, large 5 1)
Spines on chest (small 5 0, large 5 1)
Nuptial spines on fingers (small 5 0, large 5 1)
Dark bars on limbs
n (from literature)
n (examined)

SVL (mm)
Males
Females
Tympanum (hidden 5 0, distinct 5 1)
Wart (round 5 0, elongated ridge 5 1)
Belly (smooth 5 0, granular 5 1)
Belly(immaculate50, withmarbling51)
Triangular pattern between eyes

(absent 5 0, present 5 1)
Toe webbing
Vocal sac (absent 5 0, present 5 1)
Male with spines on lip margin
(absent 5 0, present 5 1)
Spinal patches on chest (small 5 0,
large 5 1)
Spines on chest (small 5 0, large 5 1)
Nuptial spines on fingers (small 5 0,
large 5 1)
Dark bars on limbs
n (from literature)
n (examined)
—
0
1
0
0
0
—
20M, 3M
—

at base
0
1
0
0
0
distinct

—
1 M, 1F

49.5–58.4
51.2–56.1
0
0–1
0
1
1
at base
1
0
0
0
1
distinct
15M, 3F
—

36.8
44.6
0
0
0
1
0
at base
0
1

0
0
0
distinct
—
1 M, 1F

Oreolalax sterlingae,
new species (Vietnam) omeimontis

47.5–56.3
55.5–59.0
0
—
0
0
0

36.8
44.6
0
0
0
1
0

Oreolalax sterlingae,
new species (Vietnam) chuanbeiensis

popei

—
20M, 8F
1M

1/4
at base
0
0
1
1
0
0
0
1
1
1
absent
distinct
20M, 20F 20M, 10F
—
—

0
0

0

—
0

1

45.1–56.3
56.0–65.7
0
0–1
0
0
0

liangbeiensis

—
20M, 4F
—

1
1

0

—
0
1

50.0–64.8
57.3–62.2
0
—
0

1
0

lichuanensis

at base
0
1
0
0
1
—
20M, 10F
1M

1/3
0
0
0
0
1
—
7M, 2F
—

57.5–73.5
62.4–70.6
1
—
0

1
0

1/4
0
1
0
0
0
—
10M, 10F
2F

at base
0
1
0
0
1
indistinct
30M, 3F
—

44.3–52.6 40.0–47.0
45.0–55.3 48.0–54.0
0
0
—
—
0

0
1
0
0
1

schmidti

—
6M, 2F
—

0
0

1

1/3–2/3
0
1

distinct
10M, 8F
—

1
1

0

—
0
—

52.6–60.0
53.3–58.2
0
0
0
0
—

—
0
—
—
—
—
distinct
1M
—

58.2
—
0
—
1
1
—

—
0
1
1
0
0
absent
10M, 10F
—

45.4–50.6
54.3–61.4
0
0
0
1
0

weigoldi xiangchengensis

—
4M
—

1
1

0

1/3

0
1

47.4–49.8
—
0
—
0
0–1
indistinct

major multipunctatus nanjiangensis
59.2–68.7
65.0–70.0
0
0
0
1
0

puxiongensis rhodostigmatus rugosus

—
16M, 4F
3M, 3F

1
1

0

1/3
0
1

48.1–60.3
48.7–56.5
0
0
0
1
1

jingdongensis

43.4–51.0 60.0–69.0 41.3–45.3
46.8–54.4 51.5–67.0 43.0–50.0
0
0
0
very small
0
1
0
0
1
0
1
0
0

0
1

pingii

—
16M, 3F
—

0
0

1

1/2–2/3
0
1

48.6–61.5
57.4–60.0
0
—
0
1
0

granulosus

Table 3. Morphological Comparison of Species of Oreolalax (M = male, F = female, — = data unavailable).

Nguyen et al.—New Oreolalax from Vietnam
217


218

Copeia 2013, No. 2

Fig. 2. Holotype (A: IEBR A.2012.1, male) and paratype (B: ZFMK 92830, female) of Oreolalax sterlingae, new species, in life.


Nguyen et al.—New Oreolalax from Vietnam

219

Fig. 4. Ventral side of hand (A) and foot (B) of the holotype (IEBR
A.2012.1) of Oreolalax sterlingae, new species.

Hindlimbs: Femur slightly longer than tibia (FeL 17.8 mm,
TbL 17.3 mm, TbL/SVL 47%); heels overlapping when held
at right angles to the body; foot long (FoL 28.3 mm); toes
long and thin, toe IV (FTL 14.5 mm) shorter than femur;
relative length of toes: I,II,V#III,IV; tip of toes rounded,
not enlarged, without discs, webbing at base only; dermal
fringe along toe V absent; subarticular tubercles indistinct;
inner metatarsal tubercle present but not developed,
rounded (IMT 2.8 mm); outer metatarsal tubercle absent;
tarsal fold absent.
Skin: Snout and dorsal head with flattened warts; dorsum
and upper flanks with distinct large warts, bearing small

black spines; dorsolateral folds absent; upper surface of
forelimbs, tarsus, and legs with flattened warts, bearing
small black spines; parotoid glands well developed with
small black spines; black spines also present in loreal region,
upper and lower jaws, tympanic region, and more distinct
on margin of lower lip; flank with white spiny spots; chest
with two distinctly separated, oval patches of small black
spines (CW 14.2 mm, SPW 4.1 mm, ratio SPW/CW 0.29,
narrowest distance between patches 3.3 mm); throat, venter,
and lower surface of limbs smooth.
Fig. 3. Ventral side (A) and chest region (B) of the holotype (IEBR
A.2012.1) of Oreolalax sterlingae, new species.

between tip of snout and eye (NS 3.2 mm, EN 3.2 mm); pupil
vertical; tympanum hidden; parotoid gland elongate (ParL
8.3/8.5 mm), about three times longer than wide; canthal,
preorbital, supraorbital, postorbital, supratympanic, and
parietal ridges absent; maxillary teeth present; vomerine
teeth absent; tongue longer than wide, free at the back,
notched posteriorly.
Forelimbs: Arm relatively short (FLL 7.4 mm), shorter than
half of hand (HAL 18.4 mm), distinctly enlarged; relative
length of fingers: I,II,IV,III, tip of finger rounded, not
enlarged, without groove; fingers without dermal fringe and
webbing; subarticular tubercles absent on fingers I and II,
indistinct on fingers III and IV; palmar tubercles flat,
rounded; inner and upper sides of fingers I and II with
small black nuptial spines.

Coloration in preservative.—Upper head grayish brown;

dorsum and upper flanks grayish brown with small black
spines; upper lip grayish brown with some indistinct whitish
spots; upper surface of arm and thigh with whitish spots or
bars; chest with two patches of black spines; lower zone of
flanks grayish brown with whitish spots; chin, throat, and
belly cream with brown mottlings; lower surface of limbs
grayish brown with whitish spots.
Coloration in life.—Dorsum and upper flanks brown with
small black spines; upper lip brown with some indistinct
whitish spots; upper surface of arm and thigh with whitish
spots or bars; lower zone of flanks brown with yellowish
white spots; chin, throat, and belly cream with brown
mottling; lower surface of limbs dark brown with whitish
spots (Figs. 2, 3, 4).
Sexual dimorphism.—Male is smaller than the female
(Table 4) and has lower lip with distinct black spines, dorsal


220

Copeia 2013, No. 2

Table 4. Measurements (in mm) of the Holotype and Paratype of
Oreolalax sterlingae, New Species. Abbreviations defined in the text.

SVL
HL
HW
SNL
NS

EN
IN
IOD
ED
UEW
DAE
DPE
MAE
MPE
MN
FLL
HAL
FFL
TFL
IPT
OPT
TbL
FoL
FeL
FTL
IMT
OMT
ParL (left/right)

IEBR A.2012.1
Holotype (Male)

ZFMK 92830
Paratype (Female)

36.8
13.3
13.7
6.1
3.2
3.2
4
4.3
5.1
3.3
6.9
11.7
9.2
5.2
11.4
7.4
18.4
4.3
8.4
1.9
2.3
17.3
28.3
17.8
14.5
2.8
absent
8.3/8.5

44.6

15.3
15.8
6.9
3.5
3.4
4.3
5.4
5.6
3.7
8.1
13.1
10.1
6.9
12.9
9.7
23.2
5.7
10.8
2.0
2.1
19.7
32.4
20.3
16.8
2.9
absent
5.5/5.5

warts more prominent, chest with two patches of small
spines, inner and upper sides of fingers I and II with black

nuptial spines. Vocal sacs are absent in both sexes.
Natural history.—Specimens of Oreolalax sterlingae were
found at night in a rocky rivulet in bamboo forest at an
elevation of 2900 m (Fig. 5). Other species of Oreolalax occur
at 700–3550 m (Wei et al., 2007; Fei et al., 2010; Frost, 2011).
The female (ZFMK 92830) contained yellow eggs (3.6 mm in
diameter).
Distribution.—Oreolalax sterlingae is known only from Hoang
Lien Mountain Range, Lao Cai Province, northern Vietnam
(Fig. 6).
Etymology.—The specific epithet is dedicated to Dr. Eleanor
J. Sterling from the Center for Biodiversity and Conservation, American Museum of Natural History in New York, to
acknowledge her outstanding contribution to biodiversity
research and conservation in Vietnam. As common names
we suggest Sterling’s Toothed Toad (English), Co´c nu´i s-tecling (Vietnamese).
DISCUSSION
Although our morphological and molecular results clearly
show that the new species belongs to the genus Oreolalax, its
position within the genus is still largely unresolved. The
sister relationship between O. sterlingae and the clade

Fig. 5. Habitat of Oreolalax sterlingae, new species, on Hoang Lien
Mountain Range, Lao Cai Province, Vietnam.

containing O. multipunctatus and other species (see Fig. 1)
is only weakly supported in the MP analysis (BP , 50%), and
the two groups become unresolved in the Bayesian analysis.
Moreover, many nodes in the genus receive low support
values in both analyses. Additional data from both mitochondrial and nuclear genes might help provide a more
robust phylogenetic hypothesis of the genus, as shown in

previous study (e.g., Fu et al., 2007).
The first discovery of a member of the genus Oreolalax
outside China once again confirms the close affinity
between northern Vietnam and southern China as shown
in other taxonomic groups (see Sterling et al., 2006). The
new record also highlights the unusual amphibian diversity
of the Hoang Lien Mountain Range, which is known as one
of the most famous regions in terms of herpetofaunal
diversity in Vietnam, and situated in one of the highest
phylogenetic diversity zones of amphibians in the world
(Fritz and Rahbek, 2012). Ohler et al. (2000) reported a total
of 42 species of amphibians from Hoang Lien Range with
descriptions of two new species, namely Leptolalax pluvialis
and Rhacophorus duboisi. Orlov et al. (2001) listed 20 species
of rhacophorids and discovered another new species of tree
frog, viz. Rhacophorus hoanglienensis. In addition, a number
of new species of amphibians have been discovered from
this region during the last decade: Leptobrachium echinatum
(Dubois and Ohler, 1998), Amolops cucae (Bain et al., 2006),
Amolops minutus and A. splendissimus (Orlov and Ho, 2007),
and Theloderma lateriticum (Bain et al., 2009).
It is important to note the rarity of this species in Vietnam.
Although numerous surveys have been conducted in Lai Chau,
Lao Cai, and Yen Bai provinces since 1998, this is the first record
of Oreolalax from the Hoang Lien Mountain Range. Because the
actual distribution of the new species is unknown, we suggest
the species should be considered Data Deficient following
IUCN’s Red List categories (IUCN, 2001). Among 17 species of
Oreolalax known from China, 13 are listed in the IUCN Red List
(2012) as near threatened to crictically endangered due to

anthropogenic threats and small distribution ranges.
MATERIAL EXAMINED
O. jingdongensis: (6) China: KIZ 91006–91007, 91011, 91155–
91157, Yunnan, Jingdong Huashan.


Nguyen et al.—New Oreolalax from Vietnam

221

LITERATURE CITED

Fig. 6. Map showing the type locality (circle) of Oreolalax sterlingae,
new species, from Hoang Lien Mountain Range, Sa Pa District, Lao Cai
Province, Vietnam.

O. liangbeiensis: (1) China: KIZ 65II0684, Sichuan, Puxiong,
Liangbei.
O. puxiongensis: (1) China: KIZ 19810703001, Sichuan,
Puxiong.
O. rugosus: (2) China: KIZ 820857, 821129, Yunnan, Yangbi.
ACKNOWLEDGMENTS
We thank the directorate of Hoang Lien National Park for
issuing relevant permits. For the loan of specimens, we are
grateful to C. Le (Hanoi), T. Nguyen (Hanoi), J. Che and J.
Wang (Kunming). Special thanks to D. Kizirian for commenting on the manuscript, E. Sterling and K. Koy for
providing the map. T. Phung thanks T. Nguyen, H. Vu, and
L. Do for assistance in the field. T. Nguyen would like to
thank M. Bonkowski for support of work in Germany and J.
Jiang for providing literature. M. Le was supported by the

National Foundation for Science and Technology Development of Vietnam (NAFOSTED: Grant No. 106.15-2010.30).
Research of T. Nguyen in Germany is funded by the
Alexander von Humboldt Foundation (VIE 1143441).

Bain, R. H., T. Q. Nguyen, and K. V. Doan. 2009. A new
species of the genus Theloderma Tschudi, 1838 (Anura:
Rhacophoridae) from northwestern Vietnam. Zootaxa
2191:58–68.
Bain, R. H., B. L. Stuart, and N. L. Orlov. 2006. Three new
Indochinese species of cascade frogs (Amphibia: Ranidae)
allied to Rana archotapnus. Copeia 2006:43–59.
Delorme, M., A. Dubois, S. Grosjean, and A. Ohler. 2006.
Une nouvelle ergotaxinomie des Megophryidae (Amphibia, Anura). Alytes 24:6–21.
Dubois, A. 1980. Notes sur la syste´matique et la re´partition
des amphibiens anoures de Chine et des regions avoisinantes IV. Classification ge´ne´rique et subge´ne´rique des
Pelobatidae Megophryinae. Bulletin Mensuel de la Socie´te´
Linne´enne de Lyon 49:469–482.
Dubois, A. 1987 ‘‘1986’’. Miscellanea taxinomica batrachologica (I). Alytes 5:7–95.
Dubois, A., and A. Ohler. 1998. A new species of
Leptobrachium (Vibrissaphora) from northern Vietnam,
with a review of the taxonomy of the genus Leptobrachium
(Pelobatidae, Megophryinae). Dumerilia 4:1–32.
Fei, L. 1999. Atlas of Amphibians of China. Henan Press of
Science and Technology.
Fei, L., S.-Q. Hu, C.-Y. Ye, and Y.-A. Huang. 2009. Fauna
Sinica. Amphibia. Volume 2. Anura. Science Press,
Chinese Academy of Science, Beijing.
Fei, L., and Y.-Z. Huang. 1983. A new species of pelobatid
toad from western Sichuan. Acta Herpetologica Sinica
2:71–75.

Fei, L., C.-Y. Ye, and H.-J. Chen. 1992. A new pelobatid toad
of genus Oreolalax (Amphibia: Anura) from YunnanGuizhou Plateau of China. Acta Biologica Plateau Sinica
6:39–44.
Fei, L., C.-Y. Ye, and J.-P. Jiang. 2010. Colored Atlas of
Chinese Amphibians. Sichuan Publishing House of Science and Technology.
Fei, L., C.-Y. Ye, and C. Li. 1999. Description of a new
pelobatid toad of genus Oreolalax–Oreolalax nanjiangensis
from Daba Mountain Region of China. Acta Zootaxonomica Sinica 24:107–113.
Fritz, S. A., and C. Rahbek. 2012. Global patterns of
amphibian phylogenetic diversity. Journal of Biogeography 39:1373–1382.
Frost, D. R. 2011. Amphibian Species of the World: an
Online Reference. Version 5.5 (31 January 2011). Electronic Database accessible at />herpetology/amphibia/index.html. American Museum of
Natural History, New York.
Fu, J., and R. W. Murphy. 1997. Phylogeny of Chinese
Oreolalax and the use of functional outgroups to select
among multiple equally parsimonious trees. Asiatic
Herpetological Research 7:38–43.
Fu, J., C. J. Weadick, and K. Bi. 2007. A phylogeny of the
high-elevation Tibetan megophryid frogs and evidence for
the multiple origins of reversed sexual size dimorphism.
Journal of Zoology 273:315–325.
Hillis, D. M., and J. J. Bull. 1993. An empirical test of
bootstrapping as a method for assessing confidence in
phylogenetic analysis. Systematic Biology 42:182–192.
Huelsenbeck, J. P., and F. Ronquist. 2001. MRBAYES:
Bayesian inference of phylogeny. Bioinformatics 17:
754–755.


222

IUCN. 2001. IUCN Red List Categories and Criteria: Version
3.1. IUCN Species Survival Commission. IUCN, Gland,
Switzerland and Cambridge, U.K.
IUCN. 2012. IUCN Red List of Threatened Species. Version
2012.1 Downloaded on 02 August 2012. www.iucnredlist.
org
Kocher, T. D., W. K. Thomas, A. Meyer, S. V. Edwards, S.
Pa
¨a
¨ bo, F. X. Villablanca, and A. C. Wilson. 1989.
Dynamics of mitochondrial DNA evolution in mammals:
amplification and sequencing with conserved primers.
Proceedings of the National Academy of Sciences of the
United States of America 86:6196–6200.
Le, M., C. J. Raxworthy, W. P. McCord, and L. Mertz.
2006. A molecular phylogeny of tortoises (Testudines:
Testudinidae) based on mitochondrial and nuclear genes.
Molecular Phylogenetic and Evolution 40:517–531.
Liu, C.-C. 1943. Two new Scutiger from Chao-Chiao-Hsien,
Sikang. Journal of the West China Border Research Society
14(B):35–39.
Liu, C.-C. 1947. Two new frogs of the genus Scutiger from
West China. Copeia 1947:123–126.
Liu, C.-C., and S.-Q. Hu. 1960. New Scutiger from China
with a discussion about the genus. Scientia Sinica
9:760–779.
Liu, C.-C., S.-Q. Hu, and L. Fei. 1979. Five new pelobatid
toads from China. Acta Zootaxonomica Sinica. Beijing
4:83–92.

Myers, G. S., and A. E. Leviton. 1962. Generic classification
of the high-altitude pelobatid toads of Asia (Scutiger,
Aelurophryne, and Oreolalax). Copeia 1962:287–291.
Ohler, A., and A. Dubois. 1992. The holotype of Megalophrys weigoldi Vogt, 1924 (Amphibia: Anura: Pelobatidae). Journal of Herpetology 26:245–249.
Ohler, A., O. Marguis, S. R. Swan, and S. Grosjean. 2000.
Amphibian biodiversity of the Hoang Lien Nature Reserve
(Lao Cai Province, northern Vietnam) with description of
two new species. Herpetozoa 13:71–87.
Orlov, N. L., and C. T. Ho. 2007. Two new species of cascade
ranids of Amolops genus (Anura: Ranidae) from Lai Chau
Province (Northwest Vietnam). Russian Journal of Herpetology 14:211–228.
Orlov, N. L., A. Lathrop, R. W. Murphy, and C. T. Ho.
2001. Frogs of the family Rhacophoridae (Anura: Amphibia) in the northern Hoang Lien Mountains (Mount Fan Si

Copeia 2013, No. 2

Pan, Sa Pa District, Lao Cai Province), Vietnam. Russian
Journal of Herpetology 8:17–44.
Palumbi, S. R., A. P. Martin, S. Romano, W. O. McMillan,
L. Stice, and G. Grabowski. 1991. The Simple Fool’s
Guide to PCR. Version 2.0. Privately published.
Posada, D., and K. A. Crandall. 1998. MODELTEST: testing
the model of DNA substitution. Bioinformatics 14:817–
818.
Sterling, E. J., M. H. Hurley, and M. D. Le. 2006. Vietnam:
A Natural History. Yale University Press, New Haven,
Connecticut.
Swofford, D. L. 2001. PAUP*: phylogenetic analysis using
parsimony (*and other methods), version 4. Sinauer
Associates, Sunderland, Massachusetts.

Thompson, J. D., T. J. Gibson, F. Plewniak, F. Jeanmougin, and D. G. Higgins. 1997. The ClustalX windows
interface: flexible strategies for multiple sequence alignment aided by quality analysis tools. Nucleic Acids
Research 25:4876–4882.
Tian, W.-S. 1983. A new species of Oreolalax—O. chuanbeiensis. Acta Herpetologica Sinica 2:59–62.
Vogt, T. 1924. Reptilien und Amphibien aus Szetschwan,
Osttibet und Tschili. Zoologischer Anzeiger 60:337–344.
Wei, G., Z.-Z. Li, N. Xu, and J.-P. Jiang. 2007. Review on the
systematics of the genus Oreolalax. Chinese Journal of
Zoology 42:169–173.
Wei, G., B. Wang, N. Xu, Z.-Z. Li, and J.-P. Jiang. 2009.
Morphological evolution from aquatic to terrestrial in the
genus Oreolalax (Amphibia, Anura, Megophryidae). Progress in Natural Science 19:1043–1048.
Wu, G.-F., E.-M. Zhao, R. F. Inger, and H. B. Shaffer. 1993.
A new frog of the genus Oreolalax (Pelobatidae) from
Sichuan, China. Journal of Herpetology 27:410–413.
Yang, D.-T., D.-S. Ma, H.-J. Chen, and F.-L. Li. 1983.
Descriptions of two new pelobatid toads from Yunnan.
Acta Zootaxonomica Sinica 8:323–327.
Zhao, E.-M., and K. Adler. 1993. Herpetology of China.
Society for the Study of Amphibians and Reptiles, Oxford,
Ohio.
Zheng, Y.-C., B.-H. Mo, Z.-J. Liu, and X.-M. Zeng. 2004.
Phylogenetic relationships of megophryid genera (Anura:
Megophryidae) based on partial sequences of mitochondrial 16S rRNA gene. Zoological Research 25:205–213.