Summary Of Biology Doctoral Thesis: Study on the diversity and distribution patterns of Amphibians (Amphibia) in karst forests of the north Vietnam and conservation measures

MINISTRY OF EDUCATION AND

VIETNAM ACADEMY

TRAINING

OF SCIENCE AND TECHNOLOGY

GRADUATE UNIVERSITY SCIENCE AND TECHNOLOGY

PHAM THE CUONG

STUDY ON THE DIVERSITY AND DISTRIBUTION PATTERNS
OF AMPHIBIANS (AMPHIBIA) IN KARST FORESTS OF THE
NORTH VIETNAM AND CONSERVATION MEASURES

Research field: Zoology
Code: 62.42.01.03

SUMMARY OF BIOLOGY DOCTORAL THESIS

HA NOI – 2018


This dissertation is submitted to the Committee of the Graduate
University of Science and Technology

Supervisors:

1. Dr. Nguyen Quang Truong
2. Dr. Le Duc Minh

Referee 1: Ass. Prof. Dr. Le Nguyen Ngat
Referee 2: Ass. Prof. Dr. Hoang Ngoc Thao
Referee 3: Dr. Hoang Van Ngoc

The research has been carried out
at the Graduate University of Science and Technology
Date of Oral Presentation:……,…../……../2018

Copy of this dissertation is available:
National Library of Viet Nam
Graduate University of Science and Technology


INTRODUCTION
1. The necessity of the research
Vietnam is recognized as one of the countries with a high level of
biodiversity in the world (Conservation International 2014). In terms of
amphibian diversity (Class Amphibia), the species number recorded
from Vietnam has been rapidly increased in recent decades: from 82
species in 1996 to 162 species in 2005 and up to 176 species in 2009
and the current species number is ca. 230 (Nguyen & Ho 1996, Nguyen
et al. 2005, 2009, Frost 2017).
Karst ecosystem is considered as a natural laboratory for taxonomic,
ecological, evolutionary and biogeographical research (Clements et al.
2006). Karst forests provide a variety of distinct microhabitats like
“isolated islands” and are noted for their high levels of endemism
(Clements et al. 2006). In Vietnam, limestone forests are extensive in
the northeastern, northwestern regions and northern Truong Son Range
(Sterling et al. 2006).

However, limestone forest is a sensitive ecosystem to the human
impacts and it is very difficult to restore. Limestone in Vietnam has
been exploited for infrastructure and economic development, for
example, quarrying for road construction, and cement industry.
Moreover, the fauna in karst forests is threatened by decline because of
forest destruction, overexploitation, tourism development and climate
change, in particular, the poikilotherm like amphibians (Sterling et al.
2006).
The research entitled "Study on the diversity and distribution patterns of
amphibians (Amphibia) in karst ecosystem of the North Vietnam and
conservation measures” aims to assess the diversity of amphibians in
limestone forests of northern Vietnam with special focus on two group
complexes of anuran frogs, Limnonectes and Odorrana, in Vietnam. In
addition, recommendations for conservation measures are also provided.
2. Objectives
- To assess the diversity of amphibians in some selected limestone
forests of northern Vietnam;
- To review taxonomic and phylogenetic relationships of two genus
complexes, Limnonectes and Odorrana, in Vietnam;
10


- To assess conservation potential and threats to the amphibian
fauna and to provide recommendations for conservation.
3. Contents
Task 1: Inventory of the species diversity of amphibians in some
karst forests in northern Vietnam: Hoa Binh Province in the
northwestern region, Cao Bang and Ha Giang provinces in the
northeastern region and Cat Ba Island, an offshore island in the Gulf of
Tonkin. This research also focuses on the discovery of new species and

new country records.
Task 2: Evaluating distribution patterns of amphibian species across
altitudinal gradient (from 0 to 1300 m), habitat types and life modes
(on the tree, on the ground, and associated with water).
Task 3: Evaluating biogeographic relationships of amphibian fauna
amongstudy sites, between mainland and offshore islands, and between
northeastern and northwestern regions to verify the evolutionary barrier
of the Red River.
Task 4: Evaluating taxonomy and phylogenetic relationships of two
genus complexes, Odorrana and Limnonectes, based on the
morphological comparison and molecular analyses.
Task 5: Assessment of conservation potential based on the following
criteria: species diversity, number of endemic and threatened species,
possibility of population restoration. Evaluation of threats to the
amphibian populations and provide recommendations for conservation
measure.
4. Scientific and conservation significances
Scientific significance: The research provides updated data about
the diversity of amphibians in the karst forests of northern Vietnam.
The research also provides novel data on taxonomy and phylogenetic
relationships of Limnonectes and Odorrana from Vietnam.
Conservation significance: The research provides baseline data for
biodiversity conservation planning and management in northern
Vietnam.
5. New findings
A total 65 species of amphibans were recorded in karst forests of the
North Vietnam, comprising 44 species recored from Ngoc Son-Ngo
11

Luong NR, 32 species recorded from Hang Kia-Pa Co NR, 33 species
recorded from Bac Me NR, 21 species recorded from Ha Lang District,
and 23 species recorded from Cat Ba NP.
New species: Three new species were described as new for science,
Odorrana mutschmanni from Cao Bang Province, Theloderma annae
and Rhacophorus hoabinhensis from Hoa Binh Province.
New records: Three species, Leptolalax minimus, Odorrana
hainanensis, and O. lipuensis, were recorded for the first time from
Vietnam and some other new provincial records from northern
Vietnam: One species from Cao Bang Province; two species from Ha
Giang Province; three species from Hai Phong city; and five species
from Hoa Binh Province.
Biogeographic relationships: Based on the list of amphibian species
recorded from northern Vietnam, the amphibian fauna of the
northeastern region was distinctly separated from that of the
northwestern region (Similarity Index = 0.6087). Moreover, the
amphibian fauna of the karst forests in mainland also differs from that
of the offshore islands (Similarity Index = 0.4706).
Distribution patterns: Most of amphibian species were recorded at
elevations below 800 m (58 species), whereas 34 species were found at
elevations above 800 m. In terms of habitat types, most of amphibian
species were recorded in the undisturbed natural forests (58 species),
while the species number recorded in the disturbed forest was 33
species and agricultural and residential areas was 16 species. The
highest number of record species were found on the ground (42
species).
In this study, we also provide taxonomic review of two poorly
known genera of amphibians, Limnonectes (8 species) and Odorrana
(25 species), based on morphological and molecular data with
descriptions of two new species for science and three new country

records for Vietnam. Phylogenetic relationships of these genera were
also discussed with remarks on some species complexes.
In addition, we evaluated the threats to the amphibian fauna and
provided some recommendations for conservation in northern Vietnam.
12


Chapter 1
LITERATURE REVIEW
1.1. Overview of amphibian research in region
Currently, a total 7.697 species of amphibians was recorded in the
world (Frost 2017). In the neigboring countries, 432 species have been
recorded from China; 152 species from Lao and 79 species from
Cambodia (Frost 2017).
In terms of conservation concern, Stuart et al. (2004) stated that
168 species of amphibian were extinction and at least 2,500 species are
under threatened in the last 20 years (1984-2004).
1.2. Overview of amphibian research in Vietnam
1.2.1. Faunistic inventory and new discoveies
Bourret (1942) reported 171 species and subspecies of amphibians
in Indochina (Vietnam, Laos, Cambodia). Nguyen and Ho (1996)
published a checklist of reptiles and amphibians of Vietnam with 82
recorded species of amphibians. Nguyen et al. (2005) listed 162 species
of amphibians. In the recent book, Nguyen et al. (2009) recorded a total
of 176 species of amphibians from Vietnam. Since 2010 more than 40
new species of amphibians have been described from Vietnam (Forst
2017).
A number of studies on amphibians were conducted in the
Northeast, Northwest, Central, Central Highlands, Southern and some
offshore islands of Vietnam.

1.2.2. Research on biology and ecology
Research on biological and ecological characters of amphibians
focused on few species that were kept in captivity, the widespread
species or endemic to Vietnam: Duttaphrynus melanostictus,
Hoplobatrachus rugulosus, Limnonectes bannaensis, Hylarana
maosonensis, Quasipaa verrucospinosa, Polypedates mutus,
Paramesotriton deloustali.
1.2.3. Research on tadpoles and bioacoustics
Descriptions of the tadpoles of the following species were
mentioned in previous studies (Hendrix et al. 2007, Gawor et a. 2009,
Wildenhues et al. 2010, Rauhaus et al. 2012, Pham et al. 2015):
Rhacophorus annamensis, Hynarana nigrovittata, Rhacophorus
13


maximus, Theloderma corticale, Amolops cremnobatus. Le Thi Quy
(2015) described the tadpoles of 21 amphibian species from Bach Ma
National Park, Thua Thien Hue Province.
In addition, the bioacoustic analysis is a useful approach to study
biology and taxonomy of amphibians. Le Trung Dung (2016) described
the biaocoustics of 11 amphibians species.
1.2.4. Research on phylogenetic relationships
Frost et al. (2006) provided a comprehensive phylogenetic tree of
amphibian species of the world. Li et al. (2008, 2009), Biju et al. (2010),
Orlov et al. (2012), Kuraishi et al. (2012), Li et al. (2012), Yu et al.
(2010, 2013), Nguyen et al. (2015), Poyarkov et al. (2015) studied on
taxonomy and phylogenetic relationships of various groups of
Rhacophoridae.
1.2.5. Research on pathology and conservation issues
Rowley et al. (2013), Nguyen et al. (2013), Martel et al. (2014),

Laking et al. (2017) and Nguyen et al. (2017) have identified a number
of amphibian populations that infected with Batrachochytrium
dendrobatidis and B. salamandrivorans.
Rowley et al. (2010, 2016) assessed threats to the populations of
amphibians in Southeast Asia, including Vietnam. Key impacts include
habitat loss, overexploitation for food, medicinal and pet trade.
1.2.6. Research on amphibians in northern Vietnam
Although numerous field excursions have been conducted in
northern Vietnam, most of surveys were conducted in evergreen forests
such as Tay Yen Tu, Tay Con Linh, Phia Oac-Phia Den, Hoang Lien,
Muong Nhe, Sop Cop, and Copia nature reserves (Hecht et al. 2013,
Bain & Nguyen 2004, Nguyen Thien Tao 2009, Orlov et al. 2001, Le
Trung Dung 2016, Pham Van Anh 2016). Research in limestone karst
ecosystems are still limited with descriptions of new species, namely
Tylototrion ziegleri from Cao Bang and Ha Giang provinces, Liuixalus
calcarius and Philautus catbaensis from Cat Ba Island (Nishikawa et
al. 2013, Milto et al. 2013).
1.2.7. Research on Limnonectes and Odorrana in Vietnam
Genus Limnonectes: Limnonectes is one of the most diverse
groups of amphibians to date with 68 currently recognized species and
14


16 of which have been described in the last ten years (Frost 2017).
According to Nguyen et al. (2009), five species of the genus
Limnonectes are currently recorded from Vietnam. However, some
taxonomic changes have been published in recent years. Due to their
morphological similarity, species that were previously thought to be
widespread are now being recognized as complexes of species with
much narrower distributions

Genus Odorrana: Odorrana is one of the most diverse groups of
amphibians to date with 58 currently recognized species, more than 20
of which have been described in the last ten years (Frost 2017). Due to
morphological similarity in the genus, species that were formally
thought to be widespread are now being recognized as complexes of
species with much narrower distributions. Further study on taxonomic
and genetic relationships should be continued.
1.3. Natural conditions of the study area
1.3.1. Natural conditions of North Vietnam
The North Vietnam bordered by China’s Guangxi Province to the
northeast and Yunnan Province to the northwest, and by Laos along its
western border (Sterling et al. 2006).
Limestone kart forests: Most of the limestone mountain area in
Vietnam is distributed in northern Viet Nam, partly in Quang Binh
province. In the Gulf of Tonkin, there are thousands of limestone islands
in Ha Long Bay next to each other with Cat Ba Island as the center
(Sterling et al. 2006).
Vegetation: The dominant forest type in northern Vietnam is
evergreen, including both broad-leaved and coniferous plants, with
pockets of semi-evergreen forest. Coastal mangrove formations and
forests growing on limestone are also important elements of the north’s
natural landscape diversity (Sterling et al. 2006).
Fauna: Northern Vietnam’s animal communities are a mixture of
tropical and subtropical species, many of which are unique to the
northern third of the country. Within the region, the Red River may be
an effective barrier to movement (or coincide with one) for some
groups, such as amphibians and reptiles, resulting in different species
and communities in the northeast and northwest (Sterling et al. 2006).
15

1.3.2. Natural conditions of survey sites
Field surveys were conducted in the limestone forestin northern
Vietnam.
Northwest:
Ngoc Son-Ngo Luong Nature Reserve (Hoa Binh Province):
This NR is located in Tan Lac and Lac Son districts with a total area of
19.254 ha. The area is characterized by limestone mountains, with steep
terrain and rugged terrain. Between the limestone mountains there are
several flat valleys running in the north-west direction. The altitude
ranges from 100-1.065 m (Birdlife International 2004).
Hang Kia-Pa Co Nature Reserve (Hoa Binh Province): Hang
Kia-Pa Co Nature Reserve is located in Mai Chau District with a total
area of 5.257 ha. There are many high and pointed limestone blocks,
the highest peak of which is 1.536m in the Northwest of the area, the
height decreasing to the East. Most of the nature reserve stands at over
500 m (Birdlife International 2004).
Northeast:
Bac Me Nature Reserve (Ha Giang Province): This NR is located
in Bac Me District with a total area of 9.042 ha,. Bac Me Nature
Reserve is located in the depression of the Gam River to the south-east
of Ha Giang Province, with steep terrain and strong splinters with many
peaks above 1000 m along the Song Gam gulf (Biodiversity Report Bac
Me Nature Reserve 2009).
Ha Lang Forest (Cao Bang Province): Ha Lang District has a
limestone karst topography that occupies most of the area, at an altitude
of 100-750 m above sea level. According to the biodiversity conservation
planning of Cao Bang Province, a nature reserve will be established in
this district with an area of 7.343 ha (Decree No. 697/QĐ-UBND).
Cat Ba National Park: Cat Ba is the largest island of the Cat Ba

Archipelago with a total area of 26.241 ha. Cat Ba National Park is
located in the limestone archipelago of hundreds of large and small
islands at the height of 100-150 m, the highest point is Cao Vong peak
331m and the area of this protected area is 16.196 ha
(www.vuonquocgiacatba.com.vn).
16


CHAPTER 2. MATERIALS AND METHODS
2.1. Field survey and study sites
Fifteen field surveys with 119 days were conducted from 2014 to 2017.
Survey sites: Cat Ba National Park (Hai Phong City), Ha Lang
Disstrict (Cao Bang Province), Bac Me Nature Reserve (Ha Giang
Province), Hang Kia-Pa Co and Ngoc Son-Ngo Luong nature reserves
(Hoa Binh Province).
Sampling of Limnonectes and Odorrana: Twenty field surveys with
200 days were conducted in more than twenty provinces in Vietnam
from 2014 to 2017.
Molecular analysis was carried out at the Institute of Ecology and
Biological Resources and Vietnam National Museum of Nature, and
Hanoi University of Science.
2.2. Methods
2.2.1. Sampling
Excursions were carried out during the day and at night in different
habitat types.
Survey transects were set up along the streams, pools, small ponds
or along the forest paths, under the leaf litter or on the tree branches, we
thus also survey the forest near the cave entrances and cliffs.
2.2.2 Laboratory work
2.2.2.1. Morphological examination

Taxonomic identification: Morphological comparisons were made
with the voucher specimens which are deposited in the collections of
the Institute of Ecology and Biological Resources and Vietnam
National Museum of Nature. For taxonomic identification, we used the
following documents: Bourret (1942), Bain et al. (2003), Ohler et al.
(2011), Orlov et al. (2003, 2006, 2012), Suwannapoom et al. (2016),
Taylor (1962), and othe related documents that were cited in each
species account. Species names followed Nguyen et al. (2009), Frost
(2017) and recently published papers.
2.2.2.2. Molecular analysis
A fragment of mitochondrial gene (cytochrome b, 12S, 16S) was
sequenced to investigate phylogenetic relationships of all species of the
genus Limnonectes and Odorrana in Vietnam.
17


The sequences were aligned using ClustalX v2.1 (Thompson et al.
1997), selected the optimal model using Modeltest v3.7 (Posada and
Crandal 1998), analyzed using Bayesian inference (BI) as implemented
in Mr.Bayes v3.1.2 (Ronquist et al. 2012).
We used BI and maximum-likelihood (ML) approaches to
reconstruct the mitochondrial gene tree (matrilineal genealogy). The
analyses used a concatenation of COI and 16S rRNA sequences.
2.2.3. Distribution patterns
Habitat: Species composition were assessed in different habitat
types (agricultural and residential areas, disturbed natural forests,
undisturbed natural forests).
Altitudinal gradient: We compared the species composition of
amphibians between elevations below 800 m and above 800 m.
Life modes: Species composition of amphibians was evaluated

based on three life modes: on the trees, on the ground, and associated
with water.
2.2.4. Conservation status
Conservation status of threatened species was referred to the Vietnam
Red Data Book (2007); IUCN Red List (2017); CITES appendices
(2017); the Governmental Decree No. 32/2006/ND-CP; and the
Governmental Decree No. 160/2013/ND-CP. The endemic species of
Vietnam are species that currenly known only from Vietnam.
2.2.5. Statistic analysis
Statistic analyses were performed with the softwware PAST
Statistics (Hammer et al. 2001)
2.2.6. Conservation issues
- Evaluation of threats to amphibians, for example habitat loss and
degradation and overharvesting.
- Providing recommendations for conservation with the following
aspects:
habitat
protection
and
restoration,
control
of
hunting/overharevesting, development of breeding program, and awareness
campaign.
2.3. Materials
A total of 524 specimens of amphibians were collected in karst
forests of the North Vietnam for morphorlogical analysis.
18

A total of 148 specimens of Limnonectes and 232 specimens of
Odorrana were collected in Vietnam for morphorlogical analysis.
A total of 150 tissue samples, consisting of 46 tissue samples of
Limnonectes, 74 tissue samples of Odorrana, and 30 tissue samples of
Liuixalus, Rhacophus, and Theloderma, were use for molecular analysis.
Chapter 3
RESULTS AND DISCUSSION
3.1. Species composition of amphibians
3.1.1. List of recorded species
A collection of 524 specimens of amphibians was collected from
different survey sites in the research areas.
Species composition: A total of 65 species belonging to 30 genera, 8
families, 3 orders, comprising 44 species (26 genera, 7 families, 2 orders)
recorded from Ngoc Son-Ngo Luong NR, 32 species (21 genera, 6 families, 1
orders) recorded from Hang Kia-Pa Co NR, 33 species (20 genera, 7 families,
2 orders) recorded from Bac Me NR, 21 species (12 genera, 5 families, 1
orders) recorded from Ha Lang District, and 23 species (15 genera, 5 families,
1 orders) recorded from Cat Ba NP (Table 3.1)
In terms of species diversity, Rhacophoridae is the most diverse
family with 9 recorded genera and 21 recorded species (Figure 3.1).

Figure 3.1. Species diversity of amphibian families

19


Table 3.1. List of amphibians recorded in the research areas
N
o

1.
2.
3.
4.
5.
6.

7.
8.
9.
10.
11.
12.
13.
14.
15.

Vietnamese name
Bộ Không đuôi
Họ Cóc
Cóc nhà
Cóc rừng
Họ Cóc bùn
Cóc mày sa pa
Cóc mày nhỏ
Cóc mày
Cóc mày đêm

Cóc núi
Cóc núi miệng nhỏ

Cóc mắt bên
Họ Nhái bầu
Cóc đốm
Ếch ương thường
Nhái bầu bec-mo
Nhái bầu bút-lơ
Nhái bầu hoa
Nhái bầu

16. Nhái bầu hây-môn
17. Nhái bầu vân
18. Nhái bầu trơn
Họ Ếch nhái
chính thức
19. Ngoé
20. Ếch đồng
21. Ếch nhẽo ban-na
22. Ếch lim-boc
23. Ếch vạch
24. Ếch gai bau-len-go
25. Ếch gai sần
26. Cóc nước sần
27. Cóc nước mac-ten
Họ Ếch nhái
28. Ếch bám đá
29. Chàng hiu
30. Chàng đài bắc
31. Ếch suối
32. Ếch xanh
33. Ếch bám đá hoa

34. Ếch hải nam
35. Ếch li-pu
36. Ếch đá mut-x-man
37.
38.
39.
40.
41.

Ếch mõm dài
Ếch ti-an-nan
Hiu hiu
Chẫu chuộc
Chàng mẫu sơn

Scientific name
Anura
Bufonidae Gray, 1825
Duttaphrynus melanostictus (Schneider, 1799)
Ingerophrynus galeatus (Günther, 1864)
Megophryidae Bonaparte, 1850
Leptobrachium chapaense (Bourret, 1937)
Leptolalax minimus (Taylor, 1962) (**)
Leptolalax pelodytoides (Boulenger, 1893)
Leptolalax nyx Ohler, Wollenberg,
Grosjean, Hendrix, Vences, Ziegler &
Dubois, 2011
Ophryophryne pachyproctus Kou, 1985
Ophyryophryne microstoma Boulenger, 1903
Megophrys major (Boulenger, 1908)

Microhylidae Günther, 1858
Kalophrynus interlineatus (Blyth, 1855)
Kaloula pulchra Gray, 1831
Microhyla berdmorei (Blyth, 1856)
Microhyla butleri Boulenger, 1900
Microhyla fissipes Boulenger, 1884(*)
Microhyla mukhlesuri Hasan, Islam,
Kuramoto, Kurabayashi & Sumida, 2014
Microhyla heymonsi Vogt, 1911
Microhyla pulchra (Hallowell, 1861)
Micryletta inornata (Boulenger, 1890)
Dicroglossidae Anderson, 1871
Fejervarya limnocharis (Gravenhost, 1829)
Hoplobatrachus rugulosus (Wiegmann, 1834)
Limnonectes bannaensis Ye, Fei & Jiang, 2007
Limnonectes limborgi (Sclater, 1892)(*)
Quasipaa delacouri (Angel, 1928)
Quasipaa boulengeri (Gunther,1899)
Quasipaa verrucospinosa (Bourret, 1937)
Occidozyga lima (Gravenhorst, 1829)
Occidozyga martensii (Peters, 1867)
Ranidae Rafinesque, 1814
Amolops ricketti (Boulenger, 1899)
Hylarana macrodactyla (Günther, 1858)(*)
Hylarana taipehensis (Van Denburgh, 1909)
Hylarana sp.
Odorrana chloronota (Günther, 1876)
Odorrana geminata Bain, Stuart, Nguyen,
Che & Rao, 2009
Odorrana hainanensis Fei, Ye & Li, 2001(**)

Odorrana lipuensis Mo, Chen, Wu, Zhang
& Zhou, 2015(**)
Odorrana mutschmanni Pham, Nguyen, Le,
Bonkowski & Ziegler, 2016(***)
Odorrana nasica (Boulenger, 1903)
Odorrana tiannanensis (Yang & Li, 1980)
Rana johnsi Smith, 1921
Sylvirana guentheri (Boulenger, 1882)
Sylvirana maosonensis (Bourret, 1937)

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1

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Study sites
2 3 4

5

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42. Ếch suối meng-la
Họ Ếch cây
43. Nhái cây sọc
44. Nhái cây wa-za
45. Nhái cây quang
46. Nhái cây nhỏ đá vôi
47. Nhái cây tay-lo
48. Nhái cây cát bà
49.
50.
51.
52.
53.
54.
55.

Ếch cây đầu to
Ếch cây mi-an-ma

Nhái cây tí hon
Ếch cây xanh đốm
Ếch cây phê
Ếch cây ki-ô
Ếch cây hòa bình

56.
57.
58.
59.

Ếch cây lớn
Ếch cây oóc-lốp
Ếch cây màng bơi đỏ
Ếch cây sần an-na

60.
61.
62.
63.

Ếch cây sần đốm trắng
Ếch cây sần bắc bộ
Ếch cây sần go-don
Ếch cây sần đỏ

Bộ Có đuôi
Họ cá cóc
64. Cá cóc zig-lơ
Bộ Không chân

Họ Ếch giun
65. Ếch giun ban-na

Sylvirana menglaensis (Fei, Ye & Xie, 2008)(*)
Rhacophoridae Hoffman, 1932
Feihyla vittata (Boulenger, 1887)
Gracixalus waza Nguyen, Le, Pham, Nguyen,
Bonkowski & Ziegler, “2012” 2013
Gracixalus quangi Rowley, Dau, Nguyen,
Cao & Nguyen, 2011(*)
Liuixalus calcarius Milto, Poyarkov, Orlov
& Nguyen, 2013
Kurixalus bisacculus (Taylor, 1962)
Philautus catbaensis Milto, Poyarkov,
Orlov & Nguyen, 2013
Polypedates megacephalus Hallowell, 1861
Polypedates mutus (Smith, 1940)
Raorchestes parvulus (Boulenger, 1893)
Rhacophorus dennysi Blanford, 1881
Rhacophorus feae (Boulenger, 1893)
Rhacophorus kio Ohler & Delorme, 2006
Rhacophorus hoabinhensis Nguyen, Pham,
Nguyen, Eto & Ziegler, 2017(***)
Rhacophorus maximus Günther, 1858(*)
Rhacophorus orlovi Ziegler & Kohler, 2001
Rhacophorus rhodopus Liu & Hu, 1962(*)
Thelodema annae Nguyen, Pham, Nguyen,
Ngo & Ziegler, 2016 (***)
Theloderma albopunctatum (Liu & Hu, 1962)
Theloderma corticale (Boulenger, 1903)(*)

Theloderma gordoni Taylor, 1962(*)
Theloderma lateriticum Bain, Nguyen & Doan,
2009(*)
Caudata Fischer von Waldheim, 1813
Salamandridae Goldfuss, 1820
Tylototriton ziegleri Nishikawa, Matsui &
Nguyen, 2013
Gymnophiona Müller, 1831
Ichthyophiidae Taylor, 1968
Ichthyophis bannanicus Yang, 1984
Total

+
+

+
+

+
+

+

+
+

+

+

+

+

+
+

+
+
+

+
+
+

+
+

+
+

+
+

+

+

+

+

+
+

+
+

+
+

+

+

+

+
+

+

+

+
+

+
+

+
+

+
+

2
1

2
3

+
+

+

+
4
4

3
2

3
3

Notes: (*)-New record for provinces, (**)-New record for Vietnam, (***)-New species. Study sites: 1Ngoc Son-Ngo Luong NR, 2-Hang Kia-Pa Co NR, 3-Bac Me NR, 4-Ha Lang District, 5-Cat Ba NP.

3.1.2. New discoveries

- New species: Three new species were described as new for
science: Odorrana mutschmanni from Cao Bang Province, Theloderma
annae, Rhacophorus hoabinhensis from Hoa Binh Province.
- New records for Vietnam: Three species were recorded for the first
time from Vietnam: Leptolalax minimus from Hoa Binh Province,
Odorrana hainanensis and O. lipuensis from Cao Bang Province.
- New provincial records: One species (Rhacophorus maximus) was
recorded for the first time from Cao Bang Province, two species
(Leptolalax minimus, Rhacophurus rhodopus) were recorded for first
21


time from Ha Giang Province, three species (Microhyla fissipes,
Hylarana macrodactyla, Theloderma corticale) were recorded for first
time from Hai Phong City, five species (Limnonectes linborgi,
Sylvirana menglaensis, Gracixalus quangi, Theloderma lateriticum, T.
gordoni) were recorded for first time Hoa Binh Province.
3.1.3. Taxonomic accounts
In this section, we described the morphological character of 40 species
based on novel data obtained from examined specimens. Below in an
example of the species description.
Sylvirana menglaensis (Fei, Ye & Xie, 2008)
Specimens examined (n=5): Three adult males IEBR 3924, 3925, 3927
(HB 2014.190, 198, HB 2015.70) and two adult females IEBR 3926, 3928
(HB 2014.199, HB 2015.28) collected from Hoa Binh Province.
Morpholigical characters: The specimens agreed well with the
description of Fei et al. (2008), Le et al. (2014). Males slightly smaller than
females (SVL 40-50 mm), head longer than wide, tympanum distinct, about
2/3 half of eye diameter, males with vocal sacs. Webbing formula I0–1II1/3–
1III1/2–1IV1–0V. Skins: Dorsum slightly shagreened, dorsolateral fold

present, ventral surface smooth. Coloration in life: Dorsum brown, flanks
grey-yellow with some large black spots, belly white.
Ecological notes: The specimens were found between 19:00 and 22:00, on
the bank of rocky stram. The surrounding habitat was mixed secondary karst
forest, composed of large, medium and small hardwoods, shrubs and vines.
Distribution: Ngoc Son-Ngo Luong and Hang Kia-Pa Co nature
reserves, Hoa Binh Province.
3.2. Comparison of species composition of amphibians
3.2.1. Between study sites
The highest Similarity Index is between Ngoc Son-Ngo Luong NR
and Hang Kia-Pa Co NRs (Similarity Index = 0.7733), followed by that
between Bac Me NR and Ha Lang District (Similarity Index = 0.6038).
The species compostion of the amphibian fauna of Cat Ba NP was
distinctly separated from those of other sites (Figure 3.2).

22


Figure 3.2. Cluster analysis of the species composition of the amphibians between
study sites (bootstrap value: 1000 replications).
Notes: NS-NL = Ngoc Son-Ngo Luong NR, HK-PC = Hang Kia-Pa Co NR, BM = Bac Me NR,
HL = Ha Lang District, CB = Cat Ba NP.

3.2.2. Between mainland and Cat Ba island
The amphibian fauna of the sites in mainland was distinctly
separated from that of Cat Ba island (Similarity Index = 0.4706).
3.2.3. Between northeastern and northwestern regions
The species composition of amphibians between the northeastern
and northwestern regions is disctinct from each other (Similarity Index
= 0.6087). This is also supported by the distinctness in morphological

characteristics and genetic distance of some species groups found on
each side of the Red River
3.2.4. Between forest areas in northeastern and northwestern

Figure 3.3. Cluster analysis of the species composition of the amphibians between forest
areas in northwestern (bootstrap value: 1000 replications)

23


Figure 3.4. Cluster analysis of the species composition of the amphibians between forest areas
in northeastern (bootstrap value: 1000 replications)

Figures 3.3 and 3.4 showed the separation of species composition
between limestone forest and evergreen forest in the northwestern and
northeastern regions.
3.3. Distribution patterns of amphibians
3.3.1. Altitudinal gradient

Figure 3.5. Species composition according altitudinal gradient

Most of amphibian species were recorded at elevations below 800 m
(58 species), whereas 34 species were found at elevations above 800 m
(Figure 3.5).
3.3.2. Habitat types
In terms of habitat types, most of amphibian species were recorded
in the undisturbed natural forests (58 species), while the species number
24

recorded in the disturbed forest was 33 species and agricultural and
residential areas was 16 species.
3.3.3. Life modes
The highest number of record species were found on the ground (42
species), 25 species found on the trees, and 22 found in associated with water.
3.4. Species diversity and phylogenetic relationships of Limnonectes
and Odorrana from Vietnam
3.4.1. Species diversity and phylogenetic relationships of the genus
Limnonectes from Vietnam
3.4.1.1. Species diversity
Eight species of Limnonectes were recorded from Vietnam (Table
3.5). One species was described as new for science, Limnonectes
quangninhensis, from Quang Ninh Province.
New provincial records: Limnonectes dabanus was recorded for the
first time from Binh Thuan, Ninh Thuan, Khanh Hoa provinces;
L. gyldenstolpei was recorded for the first time from Dong Nai
Province; L. limgorgi was recorded for the first time from Quang Ninh,
Hoa Binh, Kon Tum; L. nguyenorum was recorded for the first time
from Vinh Phuc, Son La; L. poilani was recorded for the first time from
Binh Thuan, Ninh Thuan, Khanh Hoa.
Two species, Limnonectes nguyenorum and L. quangninhensis are
currently known only from Vietnam.
Table 3.5. List of the genus Limnonectes recorded from Vietnam
No
1.
2.
3.
4.
5.
6.

7.
8.

Scientific name
Limnonectes bannaensis Ye, Fei & Jiang, 2007
L. dabanus (Smith, 1922) (*)
L. gyldenstolpei (Andersson, 1916) (*)
L. khammonensis (Smith, 1929)
L. limborgi (Sclater, 1892) (*)
L. nguyenorum Mcleod, Kurlbaum &
Hoang, 2015(*)
L. poilani (Bourret, 1942) (*)
L. quangninhensis Pham, Le, Nguyen,
Ziegler, Wu & Nguyen, 2017(**)

Vietnamese name
Ếch nhẽo ban-na
Ếch gáy dô
Ếch gyl-den-s-tol-pe
Ếch nhẽo khăm muộn
Ếch lim-boc
Ếch nhẽo nguyễn

Endemic

+

Ếch nhẽo poi-lan
Ếch nhẽo quảng ninh

+

Ghi chú: (*)-New provincial records, (**)-New species

3.4.1.2. Taxonomic accounts
We described 7 species of Odorrana recorded from Vietnam,
including additional data about distribution and ecological
characteristics.
25


A key to 8 species of the genus Limnonectes from Vietnam was also
provided.
3.4.1.3. Phylogenetic relationships
Species of genus Limnonectes from Vietnam are placed in six
independent clades, with strong support values: the Clade Icontaining
L. dabanus, L. gyldenstolpei, L. hascheanus, L. lauhachindai, L.
limborgi, L. macrognathus, and L. kohchangae; the clade IIconsisting
of L. sp1., L. sp2., and L. fragilis; the clade III containing only L.
poilani; the clade IV consisting of L. isanensis, L. jarujini, L.
megastomias, L. longchuanensis, L. nguyenorum, L. sp4., L. taylori;
the clade V containing only L. bannaensis; and the clade VI conssisting
of L. fujianensis and L. quangninhensis (Figure 3.6).

Figure 3.6. Phylogram based on Bayesian analysis of the genus Limnonectes in
Vietnam. Number above branches are Bayesian posterior probabilities

26

3.4.2. Species diversity and phylogenetic relationships of the genus
Odorrana from Vietnam
3.4.2.1. Species diversity
A total of 25 species of genus Odorrana was recorded from Vietnam
(Table 3.12). One new species was described as new for science:
Odorrana mutschmanni from Cao Bang Province; three species were
recorded for the first time from Vietnam: Odorrana hainanensis, O.
lipuensis, and O. versabilis. In addition, we provide the first descriptions
of females of two species: Oodorran gigatympana and O. trankieni.
New provincial records: Odorrana bacboensis was recorded for the
first time from Ha Tinh; O. gigatympana was recorded for the first time
from Thua Thien-Hue, Quang nam, Gia Lai, and Phu Yen provinces;
O. jingdongensis was recorded for the first time from Lai Chau and
Dien Bien provinces; O. khalam was recorded for the first time from
Quang Nam Province; O. morafkai was recorded for the first time from
Thanh Hoa, Kon Tum, and Lam Dong provinces; O. tiannanensis was
recorded for the first time from Cao Bang and Thanh Hoa provinces; O.
trankieni was recorded for the first time from Hoa Binh and Bac Giang
provinces; and O. yentuensis was recorded for the first time from
Quang Ninh Province.
Among 25 recorded species of Odorrana from Vietnam, 11 species are
globally or nationally threatened at different levels (Table 3.12).
Table 3.12. List of the genus Odorrna recorded from Vietnam
No

Scientific name

Vietnamese name

1.

2.

Odorrana absita (Stuart & Chan-ard, 2005)
O. andersonii (Boulenger, 1882)
O. bacboensis (Bain, Lathrop,
Murphy, Orlov & Ho, 2003)(*)
O. banaorum (Bain, Lathrop, Murphy,
Orlov & Ho, 2003)
O. chapaensis (Bourret, 1937)
O. chloronota (Günther, 1876)
O. geminata Bain, Stuart, Nguyen,
Che & Rao, 2009
O. gigatympana (Orlov, Ananjeva &
Ho, 2006) (*)
O. grahami (Boulenger, 1917)
O. graminea (Boulenger, 1900)
Odorrana hainanensis Fei, Ye & Li, 2001(**)

Ếch mõm ap-si-ta
Chàng an-dec-sơn

3.
4.
5.
6.
7.
8.
9.
10.
11.

27

IUCN
RL
2017

RBVN
2007

Ende
-mic

VU

Ếch bắc bộ
Ếch ba na
Ếch bám đá sa pa
Ếch xanh

NT

Ếch bám đá hoa

VU

Ếch màng nhĩ
khổng lồ
Ếch g-ra-ham
Ếch g-ra-mi-ne

Ếch hải nam

+
NT
VU


12.
13.
14.
15.
16.
17.
18.
19.
20.
21.
22.
23.
24.
25.

Ếch ging-đông
Ếch giun-li-an
Ếch ta-bu

O. jingdongensis Fei, Ye & Li, 2001(*)
O. junlianensis Huang, Fei &Ye, 2001
O. khalam (Stuart, Orlov & Chan-ard, 2005) (*)
O. lipuensis Mo, Chen, Wu, Zhang &

Zhou, 2015(**)
O. margaretae (Liu, 1950)
O. morafkai (Bain, Lathrop, Murphy,
Orlov & Ho, 2003) (*)
O. mutschmanni Pham, Nguyen, Le,
Bonkowski & Ziegler, 2016(***)
O. nasica (Boulenger, 1903)
O. orba (Stuart & Bain, 2005)
O. schmackeri (Boettger, 1892)
O. tiannanensis (Yang & Li, 1980) (*)
O. trankieni (Orlov, Le & Ho, 2003) (*)
O. versabilis (Liu & Hu, 1962) (**)
O. yentuensis Tran, Orlov & Nguyen, 2008(*)

VU
VU

Ếch li-pu
Ếch mac-ga-ret
Ếch mo-rap-kai
Ếch đá mut-x-man
Ếch mõm dài
Ếch mồ côi
Ếch s-mac-ko
Ếch ti-an-nan
Ếch trần kiên
Ếch đá
Ếch yên tử

+

+
EN

Notes. (*)-New provincial records, (**)-New records for Vietnam, (***)-New species. RBVN (2007)
= Vietnam Red Data Book (2007). Part I. Animals: VU = Vulnerabale; IUCN (2017) = IUCN Red List of
Threatened Species: EN = Endangered, VU = Vulnerabale, NT = Near Threatened.

3.4.2.2. Taxonomic accounts
We described 18 species of Odorrana recorded from Vietnam,
including additional data about distribution and ecological
characteristics.
A key to 25 species of the genus Odorrana from Vietnam was also
provided.
3.4.2.3. Phylogenetic relationships
Species of the genus Odorrana from Vietnam are placed in four
indipendent clades: the clade I consisting of O. banaorum,
O. chloronota, O. graminea, O. leporipes, O. morafkai, O. sp1, O. sp2,
O. tiannanensis, O. nasica, O. yentuensis, O. trankieni, O. nasuta,
O. bacboensis O. fengkaiensis, O. hainanensis, O. gigatympana, and
O. sp3; the clade II containing O. chapaensis and O. geminata; the
clade III consisting of O. andersonii, O. grahami, O. jingdongensis,
O. junlianensis, O. margaretae, O. mutschmanni, and O. wuchuanensis;
and the clade IV containing only O. lipuensis (Figure 3.7).

28


Hình 3.7. Phylogram based on the Bayesian analysis of the genus Odorrana in
Vietnam. Number above branches are Bayesian posterior probabilities

29


3.5. Conservation issues
3.5.1. Species of conservation concern
The amphibian fauna of karst forests of the North Vietnam
contain a high level of conservation concern with six species listed in
the Red Data Book of Vietnam (2007), six species listed in the IUCN
Red list (2017), and nine species are currently known only from Vietnam
(Table 3.16)
Table 3.16. List of threatened and endemic species
No

1.
2.
3.
4.
5.
6.
7.
8.
9.
10.
11.
12.
13.
14.
15.
16.

17.
18.
19.

Species name

Ingerophrynus galeatus
Quasipaa delacouri
Quasipaa boulengeri
Quasipaa verucospinosa
Odorrana geminate
Odorrana hainanensis
Odorrana mustchmanni
Gracixalus quangi
Gracixalus waza
Liuixalus catbaensis
Philautus catbaensis
Rhacophorus feae
Rhacophorus kio
Rhacophorus hoabinhensis
Theloderma annae
Theloderma corticale
Theloderma lateriticum
Tylototriton ziegleri
Ichthyophis bannanicus
Tổng số

RBVN
(2007)

IUCN
RL
(2017)

VU
EN

Ende
-mic

Location
NS
NL

HK
PC

BM

HL

CB

+
+
EN
NT
VU
VU

+
+
+

+
+
+
+

+
VU

+

+

+
+
+
EN
EN

+
+
+
+

+
+
EN

VU
VU
6

5

+
+
9

+
+
+

+
+
+
+

+

+

+

+

+

5

3

+
+

+
8

7

6

3.5.2. Threats to the amphibian faunas in the study sites
3.5.2.1. Habitat loss and degradation
Major causes of deforestation and degradation in the study areas
are exploitation of forest products, forest fire, timber logging, and
road construction in Hang Kia-Pa Co and Ngoc Son-Ngo Luong NRs
(Hoa Binh Province), and Bac Me NR (Ha Giang Province).
3.5.2.2. Hunting of amphibians
Amphibians were collected by local peoples for food and wildlife
trade in Ngoc Son-Ngo Luong NR (Hoa Binh Province) and Bac Me
NR (Ha Giang Province).

21


3.5.3. Conservation measures
3.5.3.1. Priority sites of conservation
Priority sites of conservation for amphibians in study areas were

evaluated on the basis of the following criteria: species richness,
number of endemic and threatened species, forest area and habitat
quality, and level of human disturbance. The Ngoc Son-Ngo Luong
NR and Cat Ba NP are two sites with a high level of conservation
priority.
3.5.3.2. Species of conservation concern
Species of conservation consern comprises all threatened and
endemic species, for instance the members of Quasipaa,
Rhacophorus, Theloderma, and Tylototriton.
3.5.3.3. Conservation activities
The following conservation activities are recommended in order to
protect the habitat and populations of amphibians in northern
Vietnam:
Establishment of the Ha Lang Nature Reserve in Cao Bang
Province.
Protecting the existing fores area in all survey sites in northern
Vietnam and linkage of isolated forest patches.
Sustainable use of amphibians for food consumption and control
hunting activities in breeding season. Protecting all endemic and
threatened species
Developing a breeding program for some amphibian species in
order to provide breeding animals for food consumption or pet trade:
Limnonectes, Quasipaa, Rhacophorus, and Theloderma.
Economic development and ecotourism: Developing a cultural
tourism program, combining nature discovery and animal observation
in Ngoc Son-Ngo Luong, Hang Kia-Pa Co, Ha Lang and on Cat Ba
Island.
Raising awareness campaign about amphibian crisis and
conservation.

22


CONCLUSIONS AND RECOMMENDATIONS
Conclusions
1. Species diversity: A total 65 species of amphibans were
recorded in karst forests of the North Vietnam, comprising 44 species
recored from Ngoc Son-Ngo Luong Nature Reserve, 32 species
recorded from Hang Kia-Pa Co Nature Reserve, 33 species recorded
from Bac Me Nature Reserve, 21 species recorded from Ha Lang
District, and 23 species recorded from Cat Ba National Park. Three
new species were described as new for science, Odorrana
mutschmanni, Theloderma annae, and Rhacophorus hoabinhensis.
Three species, Leptolalax minimus, Odorrana hainanensis, O.
lipuensis, were recorded for the first time from Vietnam and some
other new provincial records from northern Vietnam: one species
from Cao Bang Province, two species from Ha Giang Province, three
species from Hai Phong City, and five species from Hoa Binh
Province.
2. Biogeographic relationships: The amphibian fauna of the
northeastern region was distinctly separated from that of the
northwestern region (Similarity Index = 0.6087). The amphibian
fauna of the karst forests in mainland also differs from that of the
offshore islands (Similarity Index = 0.4706).
3. Distribution patterns: Most of amphibian species were
recorded at elevations below 800 m (58 species), whereas 34 species
were found at elevations above 800 m. In terms of habitat types, most
of amphibian species were recorded in the undisturbed natural forests
(58 species), while the species number recorded in the disturbed
forest was 33 species and agricultural and residential areas was 16

species. The highest number of record species were found on the
ground (42 species), 25 species were found on the trees, and 22 were
found on associated with water.
4. Eight species of genus Limnonectes and 25 species genus
Odorrana were recorded from Vietnam, based on morphological and
molecular data with descriptions of two new species for science and
three new country records for Vietnam. In terms of phylogenetic
23