Zoosyst. Evol. 100 (3) 2024, 897-911 | DOI 10.3897/zse.100.125798

Ate
BERLIN

Revalidated after having been described more than a century ago:
Calamaria berezowskii Giinther, 1896 (Squamata, Colubridae)
from Sichuan, Southwestern China

Ya-Ting Liang!*°, Zi-Dan Huang! *°, Li Ding*, Gernot Vogel®, Natalia B. Ananjeva®,
Nikolai L. Orlov®, Sheng-Chao Shi’, Zheng-Jun Wu!*?, Ze-Ning Chen!

1 Key Laboratory of Ecology of Rare and Endangered Species and Environmental Protection (Guangxi Normal University), Ministry of Education,
Guilin 541006, Guangxi, China

Guangxi Key Laboratory of Rare and Endangered Animal Ecology, Guangxi Normal University, Guilin 541006, Guangxi, China

School of Life Sciences, Guangxi Normal University, Guilin 541006, Guangxi, China

Chengdu Institute of Biology, Chinese Academy of Sciences, Chengdu 610041, Sichuan, China

Society for Southeast Asian Herpetology, D-69115 Heidelberg, Germany

Division of Herpetology, Zoological Institute, Russian Academy of Sciences, St. Petersburg 199034, Russia

N DO oO fF W PY

Hubei Engineering Research Center for Protection and Utilization of Special Biological Resources in the Hanjiang River Basin, School of Life
Science, Jianghan University, Wuhan 430056, China

https://zoobank. org/453CA 1 DC-07E7-40E8-9979-243C9B9I48373

Corresponding authors: Natalia B. Ananjeva (nananjeva09@gmail.com); Ze-Ning Chen (chenzn@gxnu.edu.cn)

Academic editor: Justin Bernstein # Received 19 April 2024 # Accepted 9 June 2024 Published 5 July 2024

Abstract

The reed snakes of the genus Calamaria Boie, 1827 are one of the largest groups of Asian snakes, distributed from northeast India
to the Maluku Islands of east Indonesia. Recent research on the genus in China has revealed that the species diversity of the group
was underestimated. In this study, morphological comparisons and mitochondrial DNA analysis revealed that a junior synonym of
C. pavimentata Dumeéril, Bibron & Dumeéril, 1854 — Calamaria berezowskii Gunther, 1896 is valid, hence we redescribed and re-
covered the validity of C. berezowskii. This species can be distinguished from other congeners by the combination of the following
characters: four supralabials; one preocular; rostral shield width larger than height; mental not touching anterior chin shields; eye
diameter less than the distance from eye to mouth edge; less than 1/2 of the posterior chin shield meets in the midline; dorsal scales
reduced to six rows at tail; indistinct light ring present in the nuchal region or a more or less distinct yellowish collar. Phylogeneti-
cally, this species is sister to C. pavimentata, with significant genetic differences (0.190) on mitochondrial gene Cyt b.

Key Words

Calamaria berezowskii, Colubridae, morphology, phylogenetics, taxonomy

Introduction 1854; C. pavimentata Duméril, Bibron & Duméril, 1854;
C. lovii Boulenger, 1887; C. septentrionalis Boulenger,

Calamaria Boie, 1827 is the largest group of the colubrid 1890; C. prakkei Lidth De Jeude, 1893; C. buchi Marx &

subfamily Calamariinae (Reed Snakes), containing more
than 68 species (Uetz et al. 2023). In mainland Southeast
Asiaand China, 20 species of Calamaria are known at pres-
ent, including C. /umbricoidea Boie, 1827; C. albiventer
(Gray, 1834); C. schlegeli Duméril, Bibron & Dumeéril,

Inger, 1955; C. yunnanensis Chernov, 1962 (Yeung et al.
2022; Uetz et al. 2023); C. thanhi Ziegler & Quyet, 2005;
C. gialaiensis Ziegler, Nguyen & Nguyen, 2009; C. san-
gi Nguyen, Koch et Ziegler, 2009; C. abramovi Orlov,
2009; C. concolor Orlov, Nguyen, Nguyen, Ananjeva &

Copyright Liang, Y.-T. et al. This is an open access article distributed under the terms of the Creative Commons Attribution License (CC BY 4.0), which permits unrestricted use,
distribution, and reproduction in any medium, provided the original author and source are credited.

898

Ho, 2010; C. andersoni Yang & Zheng, 2018; C. dominici
Ziegler, Tran & Nguyen, 2019; C. strigiventris Poyarkov,
Nguyen, Orlov & Vogel, 2019; C. nebulosa Lee, 2021;
C. arcana Yeung, Lau & Yang, 2022; C. jinggangensis
Cai, Jiang, Wu, Huang, Fei & Ding, 2023.

There were only three species of genus Calamaria re-
corded in China 26 years ago: C. pavimentata, C. septen-
trionalis and C. yunnanensis (Zhao et al. 1998). Recently,
three more species were described: C. andersoni, C. ar-
cana, C. jinggangensis (Yang and Zheng 2018; Yeung et
al. 2022; Cai et al. 2023; Uetz et al. 2023). The species
diversity of the genus in China is suggested to be under-
estimated, and those widely distributed species should be
re-evaluated. For example, the widely recorded species
C. pavimentata has a synonym, C. berezowskii Gunther
1896, which was described based on two specimens from
Lun-ngan-fu (7% )ff Long’an Fu, now J 2244 Long’an
Town of */ 70 Pingwu County) of Sze-chuen (Sichuan
Province of China) (Gunther 1896).

During the scientific expeditions in the Gongga Moun-
tains, Luding County, Sichuan Province, between 2017
and 2022, we collected three Calamaria specimens. Mor-
phological and molecular phylogenetic analyses showed
that these specimens represent a species that differs from
all currently recognized congeners of the genus. However,

100°0'0"E

35°0'0"N

30°0'0"N

100°0'0"E

Liang, Y.-T. et al.: Calamaria berezowskii Gunther, 1896

the morphology of these specimens matches the original
description of C. berezowskii. Besides, Luding County
and Pingwu County both are located on the eastern slope
of Qinghai-Tibet Plateau bordering the Sichuan Basin.
Therefore, we identify the specimens as C. berezowshkii,
and the species is revalidated and redescribed here.

Materials and methods
Sampling

Three specimens of Calamaria were collected from east-
ern slope of Mt. Gongga, Mox1 Town, Luding County,
Ganzi Tibetan Autonomous Prefecture, Sichuan Province,
China during 2017 to 2022 (Fig. 1). One adult female
GXNU DLR195 (29.645105°N, 102.111076°E, 1736 m
a.s.l.), one adult male GXNU DLR194 (29.615872°N,
102.109208°E, 1680 m as.l.), one juvenile female
GXNU_ 20221215002 (29.641749°N, 102.110340°E,
1827 ma.s.l.), collected by Xu Zhang on 31 August 2018,
Li Ding on 21 August 2017 and Congcong Du on 15 De-
cember 2022. The three specimens were fixed and stored
in 80% ethanol and deposited at the School of Life Sci-
ences, Guangxi Normal University.

105°0'0"E 110°0'0"E

3 5°0'0"N

~ Low: -505

105°0'0"E 110°0'0"E

Figure 1. Distribution map of Calamaria berezowskii. Red star indicates the type locality, purple triangle indicates the distribution

found in this study.

zse.pensoft.net

Zoosyst. Evol. 100 (3) 2024, 897-911
Molecular phylogenetic analysis

Genomic DNA was extracted from muscle or liver tissue
taken from the collected specimens using the TIANamp
Marine Animals DNA Kit (TIANGEN Biotech). We
sequenced a fragment of the mitochondrial gene cyto-
chrome 5 (Cyt 5) using the primer pair L14910/H16064
(Burbrink et al. 2000). PCR amplification was per-
formed in 25 pl reactions containing 12.5 wl 2x SanTaq
PCR Master Mix (with Blue Dye), 10 pl ddH,O, 1 pl
F-primers, 1 ul R-primers and 0.5 ul DNA template. The
PCR conditions were an initial denaturing step at 94 °C
for 5 min, followed by 35 cycles: denaturing at 94 °C
for 30 s, annealing at 48.5 °C for 45 s, an extension
step at 72 °C for 35 s; and a final extension of 72 °C for
10 min. Finally, the PCR products were sent to Sangon
Biotechnologies Co., Ltd. (Shanghai, China), where the
purified PCR products were sequenced using the same
forward and reverse primers. Sequences for comparison
of available species were downloaded from GenBank
(Table 1).

We evaluated and manually corrected the sequencing
peak maps of the three sequences obtained, downloaded
29 Cyt b sequences of 12 species of the genus Calamaria
that have been reported in Southeast Asia and China in

Table 1. DNA sequences used in this study.

899

GenBank, and selected three species Elaphe quatuorlin-
eata, Lycodon rufozonatus and Orientocoluber spinalis as
outgroups (Yang and Zheng 2018; Yeung et al. 2022), for
a total of 35 sequences to be used for multiple sequence
comparisons. All sequences were aligned with other re-
trieved sequences in the same gene loci by using software
MEGA 11 (Kumar et al. 2018). Phylogenetic trees were
constructed based on mitochondrial gene Cyt 6b using
maximum likelihood (ML) and Bayesian inference (BI).
Maximum likelihood analysis was conducted in RAXxML
v8.2.4 (Stamatakis 2014). Confidence intervals were de-
termined with 1000 bootstrap replicates utilizing the rap-
id bootstrap option under the GTR+GAMMA substitu-
tion model. Bayesian inference analysis was conducted in
MrBayes 3.2 (Ronquist et al. 2012). PartitionFinder 2.1.1
software was used to select the most suitable nucleotide
substitution models for Cyt b sequence data based on the
Bayesian information criterion (BIC): GTR+G, HKY+G
and GTR+G (Lanfear et al. 2012; Lanfear et al. 2017).
We ran our analyses for 20 million generations with the
chains, sampling every 1000 generations using the Mar-
kov Chain Monte Carlo (MCMC). After removing out-
group taxa, MEGA11 (Kumar et al. 2018) was used to
calculate uncorrected pairwise sequence divergence be-
tween the Calamaria species.

No. Taxa Voucher Locality Cyt b Reference

1 C. septentrionalis HSR19100 Mt. Huangshan, Anhui, China 0Q354842 Cai et al. 2023

2 C. septentrionalis HS11145 Mt. Nanling, Guangdong, China 0Q354840 Cai et al. 2023

3 C. septentrionalis DL2021610-1 Huangsha, Guangxi, China 0Q354838 Cai et al. 2023

4 C. septentrionalis KFBG14506 Hainan Island, China MH445956 Yang and Zheng 2018
5 C. septentrionalis ROM35605 Nguyén Binh, CaoBang, Vietnam AF471081 Lawson et al. 2005
6 C. septentrionalis ROM35597 Cao Bang, Vietnam KX694890 Alencar et al. 2016
7 C. pavimentata KFBG14507 Ningming, Guangxi, China MH445957 Yang and Zheng 2018
8 C. andersoni HSR20101 Dehong, Yunnan, China 00354844 Cai et al. 2023

9 C. andersoni HSR20181 Tengchong, Yunnan, China 0Q354845 Cai et al. 2023
10 ‘C. andersoni SYSr001699 Yingjiang, Yunnan, China MH445955 Yang and Zheng 2018
11 C. yunnanensis ROM41547 Simao, Yunnan, China KX694891 Zaher et al. 2009
12. C. yunnanensis YPx503 Unknown JQ598922 Grazziotin et al. 2012
13. -C. arcana HS17082 Mt. Dawu, Guangdong, China 0Q354835 Cai et al. 2023
14. ~C. arcana KFBG14611 Mt. Dadongshan, Guangdong, China ON482335 Yeung et al. 2022
15 C. arcana GP9975 Yongxing, Hunan, China OP980549 Cai et al. 2023
16 ~=~C. arcana DLR199 Mt. Wuyi, Fujian, China 0Q354834 Cai et al. 2023
17 _ C. jinggangensis DL20200725 Mt. Jinggangshan, Jiangxi, China 0Q354830 Cai et al. 2023
18 C. jinggangensis DL20200625-2 Mt. Jinggangshan, Jiangxi, China 0Q354831 Cai et al. 2023
19 _‘C. jinggangensis DL20200625-3 Mt. Jinggangshan, Jiangxi, China 0Q354832 Cai et al. 2023
20 —C. jinggangensis DL20200625-4 Mt. Jinggangshan, Jiangxi, China 0Q354833 Cai et al. 2023
21 ~~ ‘C. muelleri RMB1283 Gowa, South Sulawesi, Indonesia MT819391 Weinell et al. 2021
22 ~~ C. muelleri TNHC58955 Gowa, South Sulawesi, Indonesia MT819390 Weinell et al. 2021
23 ~~ ‘C. lumbricoidea USMHC1560 Penang, Peninsular, Malaysia MN338526 Quah et al. 2019
24 ~ C. palavanensis KU311411 Mt. Mantalingahan, Palawan, Philippine MT819387 Weinell et al. 2021
25 C. palavanensis KU309445 Barangay lrawan, Palawan, Philippine MT819386 Weinell et al. 2021
26 ~—sC..: gervaisii KU334485 Municipality, llocos Sur, Philippines MT819385 Weinell et al. 2021
27 ~—=«C.: gervaisii KU324661 Puguis, Benguet, Philippines MT819384 Weinell et al. 2021
28 ~=-C.. schlegeli LSUHC10278 Perak, Peninsular, Malaysia MN338525 Quah et al. 2019
29. ~=C. nebulosa FMNH258666 Phongsaly, Laos MN338524 Quah et al. 2019
30 ~~‘ C.. berezowskii GXNU DLR194 Mt. Gongga, Sichuan, China PP747047 This study

31. ‘C. berezowskii GXNU DLR195 Mt. Gongga, Sichuan, China PP747048 This study

32 C. berezowskii GXNU 20221215002 Mt. Gongga, Sichuan, China PP747049 This study

33. Elaphe quatuorlineata LSUMZ40626 Turkey, European Turkey AY486931 Nagy et al. 2004
34 — Lycodon rufozonatus LSUMZ44977 Unknown AF471063 Lawson et al. 2005
35 Orientocoluber spinalis MVZ211019 Yinnan, Ningxia, China AY486924 Nagy et al. 2004

zse.pensoft.net

900
Morphological analysis

Terminology and descriptions follow the views of Inger
and Marx (1965) and Ziegler et al. (2008). Body and tail
length were measured with a tape ruler to the nearest 1 mm:
total length (TL), from the tip of snout to the tip of tail;
snout-vent length (SVL), from the tip of snout to posterior
margin of cloaca; tail length (TaL), from posterior margin
of cloaca to the tip of tail. Other measurements were con-
ducted with a digital caliper to the nearest 0.1 mm: head
length (HL), from the snout tip to the posterior margin of
the mandible; head width (HW), measured at the widest
part of the head on posterior side; head height (HH), at
the maximal highest part of the head; the eye horizontal
diameter (EyeD); and eye-mouth distance (Eye-Mouth
D), measured from the anterior point of the eye to the
mouth gap. Ventral scales (VEN) were counted according
to Dowling (1951). The enlarged shield(s) anterior to the
first ventral were regarded as preventral(s). The number of
dorsal scale rows (DSR) are given at one head length be-
hind head, at midbody, and at one head length before vent,
respectively. The tail tip was not included in the number of
subcaudal scales (SC). Sex was determined by examining
the presence or absence of hemipenis.

Type specimen (lectotype ZISP 8823) was examined
for morphological comparisons. Relevant morphological
data of other Calamaria species were obtained from ex-
amined specimens (Appendix 1) and literatures.

Institution acronyms

FMNH: Field Museum of Natural History, Chicago.
GXNU: Guangxi Normal University. KFBG: Kadoorie
Farm and Botanic Garden. KU: University of Kansas
Biodiversity Institute. LSUHC: La Sierra University

Liang, Y.-T. et al.: Calamaria berezowskii Gunther, 1896

Herpetological Collections. LSUMZ: Louisiana State
University Museum of Natural Science. MVZ: Muse-
um of Vertebrate Zoology. RMB: Rafe M. Brown field
tag (specimen deposited in Museum Zoologicum Bo-
goriense, Indonesia). ROM: Royal Ontario Museum,
Canada. SYS: Biological Museum of Sun Yat-sen Uni-
versity, Guangzhou, China. TNHC: Texas Natural Histo-
ry Collections. USMHC: Universiti Sains Malaysia Her-
petological Collection, Malaysia. ZFMK: Zoologisches
Forschungsmuseum Alexander Koenig, Bonn, Germany.
ZISP: Zoological Institute, Russian Academy of Scienc-
es, St. Petersburg, Russia.

Molecular results

The DNA dataset contains 35 samples with a total of 1105
base pairs. The phylogenetic topologies that resulted from
Bayesian Inference (BI) and Maximum likelihood analy-
sis (ML) are generally consistent in phylogenetic structure
(Fig. 2). Phylogenetically, the species Calamaria from Mt.
Gongga strongly clustered into a single lineage with high
nodal support (Fig. 2, BPP 1.00 / UFB 100). Subsequently,
it clustered with C. pavimentata KFBG14507 from Ning-
ming, Guangxi (BPP 0.96 / UFB 94), and was clustered into
the same clade with C. arcana, C. jinggangensis, C. septen-
trionalis, C. pavimentata, C. andersoni and C. yunnanensis.

The uncorrected pairwise p-distances of Cyt b sequenc-
es between the specimen of Mt. Gongga and the other 12
congeners included in the study were 17.1—31.2%, with
the minimum value observed in the comparison with
sequences of C. arcana (P = 17.1%) (Table 2). This di-
vergence is clearly among interspecies level since these
levels of divergences are distinctly higher than those ob-
served between two other well distinguished species of
C. andersoni and C. yunananensis (P = 12.2—12.6%). The

Table 2. Uncorrected p-distances between Calamaria species based on 1105 base pairs from the mitochondrial genes Cyt 6. The

serial numbers in Table 2 are consistent with those in Table 1.

No. Taxa 1-6 7 8-10 11-12 13-16
1-6 _C. septentrionalis 0.000-0.038
7 C. pavimentata 0.174-0.194
8-10 C. andersoni 0.130-0.148 0.246 0-0.019
11-12 C. yunnanensis 0.159-0.164 0.237 0.122-0.126 0.000
13-16 C. arcana 0.090-0.116 0.179- 0.181-0.206 0.183- 0.007-
0.199 0.209 0.034
17-20 C. jinggangensis 0.095-0.103 0.180 0.161-0.171 0.158 0.062-
0.074
21-22 C. muelleri 0.202-0.223 0.266 0.228-0.238 0.261- 0.178-
0.273 0.198
23 C. lumbricoidea 0.243-0.248 0.258 0.244-0.256 0.251 0.228-
0.239
24-25 C. palavanensis 0.187-0.222 0.231- 0.227-0.243 0.254- 0.207-
0.242 0.266 0.229
26-27 C. gervaisii 0.178-0.207 0.243- 0.202-0.212 0.243- 0.197-
0.258 0.244 0.223
28 C. schlegeli 0.232-0.243 0.263 0.258 0.271 0.217-
0.228
29 C. nebulosa 0.186-0.196 0.214 0.172 0.173 0.162-
0.182
30-32 C. berezowskii 0.187- 0.190 0.216- 0.208 0.171-
0.203 0.226 0.200

zse.pensoft.net

17-20. 21-22 23 24-25. 26-27 28 29 30-32
0.000
0.169- 0.007
0.178
0.228 0.164
0.173- O0.167- 0.166- 0.022
0.192 0182 0.181
0.159- 0173 0197- 0112 0.100
0.163 0.183 0.223 0.174
0.217 0.182- 0205 0170 0.186-
0.192 0.175 0.218
0.176 0.208 0.193 0.202- 0187- 0.197
0.219 0.207 0.234
0.202 0.223- 0.255 0.254 0.254 0.312 0.176 0.000
0.234 0.277. 0.271

Zoosyst. Evol. 100 (3) 2024, 897-911 901
1.00/100 Peps iane Pitted bee Atat i sapteveotss KFBG14611
aa bein | GP9975 C. arcana
1.00/99 ~ DLR199
1.00/100 eng
~ DL20200625-4
~ DL20200625-3 C. jinggangensis
1.00/100 |. ~ DL20200625-2
0.99/97 | en DL20200725
Sas ttites PSE CAAA B83 tc 8b Res UE HSR19100
0.79/79
enn HS11145 C. septentrionalis
1.00/100 OPD2 Fteteacttenbiceth DL2021610-1 x
posses ROMIZ5605
1.00/100
noi 1.00/99 poo HSR20101
con 1.00/100 ~ HSR20181 C. andersoni
1.00/100 om SYS1001699
~ ROM41547 :
1.00/100 b----- YPx503 2 ann enen sts
1.00/1 00 iajhhs piste ee GXNU DLR1 94
1.00/100 0.64/88 Pe GXNU DLR195 C. berezowskii
0.96/94 ~ GXNU 2022121500
KFBG14507 | C. pavimentata
eae FMNH258666 IC. nebulosa
10008 ers lc palavanensis
1.00/100 ~ KU309445
0.99/96 » KU334485 he
Patten le gervaisil
0.99/99 1.00/100 ~ RMB1283 .
a3 aint saoss | C. muelleri
- USMHC1560 1 C. lumbricoidea
~ LSUHC10278 IC. schlegeli
0.96 ooo LSUMZ40626 | Elaphe quatuorlineata
VUES OYSTELOT GPa | eee | EE ethics Pe tale CELL OPS nw Lk Bierseln Oe LSUM/744977 | Lycodon rufozonatus
~ MVZ211019 | Orientocoluber spinalis

Figure 2. Phylogenetic relationships of Calamaria species based on Cyt 5. Bayesian posterior probabilities (BPP) and Ultrafast
bootstrap approximation (UFB) were denoted beside each node (those lower than 75/0.7 were denoted as “-”).

Calamaria specimens from Mt. Gongga, Luding, Sich-
uan represent an evolutionary distinct lineage.

Morphological results

Morphology of the three Calamaria specimens from
Luding County, Sichuan province matches the original
description of C. berezowskii Gunther 1896 and the mor-
phological characters of lectotype ZISP 8823 (Figs 3, 4,
5, 6). They share the following characters: one preocular
and one postocular; presence of a more or less distinct
yellowish collar; similar number of ventral scales (149-—
165 vs 155-167) and subcaudal scales (16—25 vs 14—22);
presence of a faint dark line on each side of the back; tail
blunt and conical at the end. Besides, Luding County and
the type locality Pingwu County are both located on the

eastern slope of Qinghai-Tibet Plateau bordering the Si-
chuan Basin, with a geographic distance of about 350 km
(Fig. 1). Hence, the Luding specimens were identified as
conspecific with C. berezowskii.

Morphological comparisons based on some characters
between Calamaria berezowskii and other known species
of the genus from mainland Southeast Asia and China
were listed in Table 3. It can be distinguished by a com-
bination of the following characters: four supralabials;
preocular present; rostral shield width greater than height;
mental not touching anterior chin shields; eye diameter
less than distance from eye to mouth edge; less than 1/2
of the posterior chin shield meets in the midline; dorsal
scales reduced to six rows at tail; presence of indistinct
light rings in the nuchal region or a more or less distinct
yellowish collar; and the two outermost dorsal scale rows
light khaki, with upper margins partly dark pigmented.

zse.pensoft.net

902

Liang, Y.-T. et al.: Calamaria berezowskii Gunther, 1896

Table 3. Morphological comparisons of species of Calamaria from mainland Southeast Asia and China. “1+***” indicates preven-

trals + ventrals. Entries marked with ‘—’ are not available.

Species

and width
Supralabials
End of tail

paraparietal
only in anteriorly (0)

and eye-mouth distance
from (0) anterior chin shields

Comparison between eye diameter
Number of scales surrounding
Mental touching (1) or separated
Posterior chin shields meeting in
midline (1), diverging or meeting

Comparison between rostral height
Preocular: present (1) or absent (0)

C. berezowskii

C. pavimentata Sharp point

Ventrals
Subcaudals
Total length

s
=]
o
o
ea]
>
—_
0:
x
oO
=
zo}
o
=
Zo}
je)
=

=

above position of subcaudal on tail

w
co ©

=
©

(ae)
=
4 Obtuse point 24+149- | 153-— | 22-25 Not reduced | 248-290 | 123-305
155 2+165

Last-13"

Dorsal scales reducing to four rows

| Female
Male
Female

115485

subcaudal

C. septentrionalis

Broadly rounded

8-9 | 125-168 | 137-206} 3-33 | 8-20
8-9 | 148-166 | 168-188] 15-19] 6-11

Not reduced | 111-344 |117-384

C. yunnanensis H<W 4 Obtuse point | 8-9 | 167-184 15-20 3™ast 245-300 516
subcaudal

C. andersoni <W EEE Obtuse point Peper pet 2™_last 351.4 -
subcaudal

C. arcana H<W] > 1/4 Obtuse point 170-176 192 | 20-22 Not reduced | 144-303.2] 36.5

C.lumbricoidea |H<W]} = | 45]1) 5 Sharp point | 9-11 | 144-196 | 137-229] 17-27} 13-21) Last-11" 149-498 | 120-642

/H>W subcaudal

C. albiventer H<W ft 5 1 1 Sharp point Wi 147-162} 21-22 | 15-19 5th_gth 170-361
subcaudal

C. schlegeli Blunt 9-10 | 129-161 | 136-180] 25-44 | 19-37 3-25" -
subcaudal
subcaudal

C. prakkei Sharp point 7-8 | 126-132 |142-144| 31-32 | 24-25 7%] 5% 172-245 |230-256
subcaudal

C. buchi ape Obtuse point fs sgn 389-466
subcaudal

C. thanhi Gradually to a 184 198 28 Not reduced 455

point

C. gialaiensis aa Rounded He Disc aal 23 Last subcaudal -

C. sangi Obtuse point 2+190 19 Last-3" 373.3
subcaudal

C. abramovi Sharp point oie a 26 20 |Last subcaudal - 482

C. concolor Obtuse point 34+209 19 Last subcaudal 578 -

C. dominici Obtuse point cae aad - |17-18 5th_6th -
subcaudal

C. strigiventris Abruptly to point | 9-11 | 130-157 | 158-180) 29-33 | 20-30 Last-6" -
subcaudal

C. nebulosa Obtuse point 22 |Last subcaudal - 354

Taxonomy second and third supralabials entering orbit, the fourth

Calamaria berezowskii Giinther, 1896
Figs 3, 4, 5, 6

Calamaria pavimentata — populations in Sichuan of Zhao et al. (1998)
and Zhao (2006); Zhao (2003). Synonym.

Description of lectotype ZISP 8823. Adult male, col-
lected from Lun-ngan-fu (Z#)f Long’an Fu, now 7%
24H Long’an Town of */ zt Pingwu County) of Sze-
chuen (Sichuan Province of China) (Gunther 1896).

HL: 8.0 mm, 3.0% of SVL; HW: 5.0 mm, HW/HL:
62.5%; HH: 4.0 mm, HH/HL: 50.0%; EyeD: 0.7 mm, larg-
er than eye-mouth distance 1.0 mm; EyeD 8.8% of HL.

SVL: 271 mm; TaL: 19 mm; TL: 290 mm; Tal/TL: 6.6%.

DSR: 13-13-13 scales; VEN: 155; SC: 22, divided,
followed by a shield covering tail tip; anal scale single.

Rostral as broad as high; pupil rounded; preocular 1/1
(left/right, hereafter); postocular 1/1; supralabials 4/4, the

zse.pensoft.net

longest; infralabials 5/5. Visible yellowish collar on left
and right sides of the neck present. Dorsal color brown,
with a faint dark line along each side of the back; ventral
surface uniform white. Tail rather obtuse, with a conical
end. 2-3 pairs light color spots at the base of the tail.

Description of referred specimen GXNU DLR195.
Adult female, collected from Mt. Gongga, Moxi Town,
Luding County, Ganzi Tibetan Autonomous Prefecture,
Sichuan Province, China (29.645105°N, 102.111076°E,
1736 ma.s.l.) collected by Xu Zhang on 31 August 2018.

Body elongated, cylindrical; head small, not distinct
from nape; tail short, similar in form to head; tail not flat-
tened, tapering and bluntly pointed at tip.

HL: 8.3 mm, 2.9% of SVL; HW: 3.9 mm, HW/HL:
47.0%; HH: 4.0 mm, HH/HL: 48.2%; EyeD: 0.7 mm, larg-
er than eye-mouth distance 1.1 mm; EyeD 8.4% of HL.

SVL: 288 mm; TaL: 17 mm; TL: 305 mm; Tal/TL:
5.6%. Body thickness about 4.70 to 6.71 mm; base of tail
3.18 mm thick.

Zoosyst. Evol. 100 (3) 2024, 897-911

Table 3. Continued.

903

bs So/s—|8
BS = 5 2z|S2O\3
52 B = 5/2 e\as
ke S 2 YO! @ Bow
XG id - 5 8lm a] &
. = S °o E Ll— Fo ®
Species = |o0 = = Sols 5/53 References
Fa S = gals s|E 8
. 5 Bele ties
fs) = &\|Sa
eal eee 8 8 3 elZsl=
2] § 5.5/9 “5
uw Qa
C. berezowskii |6.6-10.5|5.6-6.5| — Blackish-brown or brown Light khaki or white | 0 0-1 Giinther 1896; this study
C. pavimentata |6.9-16.9] 3.7-8.5 Yellow 0 0-1 ] Inger and Marx 1965; Ziegler et al. 2008; Nguyen et al. 2009
C. septentrionalis| 6.3-8.6 | 2.6-4.3 Dark brown to black Yellow 1 1 | Inger and Marx 1965; Ziegler et al. 2008; Poyarkov et al. 2019
C. yunnanensis | 5.4-8.2 Bluish-grey or olive-brown Red or yellow 0-1} 1 Chernov 1962; Stuart and Heatwole 2008; Yang and Rao
2008; Lee 2021
C. andersoni 9.2 - Brownish Yellow 0 Yang and Zheng 2018
C. arcana 7.2-11.8| 4.7 Grey-brown with somewhat Orangish-red (in life) 1 Yeung et al. 2022; Zhang et al. 2023; Cai et al. 2023
iridescent (in life) or caramel or light yellowish-
brown (in alcohol) beige (in alcohol)
C. jinggangensis | 7.1-10.1 | 3.6-4.6 | Brownish black with iridescent (in | Dark orange (in life) 0-1 Cai et al. 2023; this study
life) or brownish black (in alcohol) | — or light khaki (in
alcohol)
C. lumbricoidea_ |6.3-11.4} 3.9-8.3 Dark brown to black Yellow 0-1 Inger and Marx 1965; Weinell et al. 2021; Lee 2021
C. albiventer 8.8-9.3 | 4.7-8.8 Brown Red 1 0 Inger and Marx 1965; Wallach et al. 2014; Lee 2021
C. schlegeli Dark brown to black White or yellow O | 0-1 Inger and Max 1965; Quah et al. 2018; Weinell et al. 2021
C. lovii gimletii Dark brown Yellow O-1 | O-1 | Inger and Marx 1965; Quah et al. 2018; Grismer et al. 2004;
Lee 2021
C. prakkei Brown Yellow 1 - Inger and Marx 1965; Wallach et al. 2014
C. buchi Black Yellow O | Inger and Marx 1965; Ziegler er al. 2008; Nguyen et al. 2009
C. thanhi Dark blue to grey Yellow 1 Ziegler and Quyet 2005; Ziegler et al. 2007; Ziegler et al.
2008; Nguyen et al. 2009; Wallach et al. 2014
C. gialaiensis 8.1 - Grey-brown Yellow 0 0 1 Ziegler et al. 2008; Wallach et al. 2014; Lee 2021
C. sangi 6.2 Greyish-brown yellow 0 1 0 Nguyen et al. 2009; Wallach et al. 2014
C. abramovi 13.3 7.1 Black Black and yellow 1 0 Orlov 2009; Wallach et al. 2014
C. concolor 7.3 - Brown Cream 0 0 Orlov et al. 2010; Wallach et al. 2014
C. dominici 5 6.2 Black Yellow and black 0 Ziegler et al. 2019
C. strigiventris 13.8- Slate grey to grey-brown Yellow and black 0 Poyarkov et al. 2019
17.9
C. nebulosa - Bluish-grey Yellow mre Lee 2021

VEN: 165 (+2 preventrals); SC: 16, all paired; anal
shield entire, ventral scales immaculate.

DSR: 13-13-13 scales, dorsal scales smooth and im-
maculate.

Rostral shield width (2.22 mm) is larger than height
(1.62 mm), internasals and prefrontals fused 2 scales;
prefrontal length (2.28 mm) is less than frontal length
(2.61 mm), not entering orbit, and touching first two su-
pralabials; frontal hexagonal, longer (2.61 mm) than wide
(2.50 mm); six paraparietals; parietal scales long, tangent
to supraocular, postocular scales, supralabials; one preoc-
ular present; parietal broadly in contact with the last supra-
labials; pupil rounded; supralabials 4/4, second and third
entering orbit, the fourth largest (length 2.34/2.32 mm)
and tangent to the postoculars; mental not touching ante-
rior chin shields; infralabials 5/5, first three touching an-
terior chin shields; anterior chin shields are slightly longer
than posterior chin shields, presence of mental groove;
less than 1/2 of the posterior chin shield meets in the mid-
line; dorsal scales reduced to 6 rows above last subcaudal
at tail; anal scale 1s complete and single.

Coloration in life. The dorsal color was blackish
brown, with a faint dark line along each side of the back
which is about 3 scales wide apart; the outermost cor-
ners of the ventral scales were brownish, and the ventral

surface was lighter; presence of distinct yellowish collar;
absence of light ring at the base of the tail.

Coloration in preservative. The specimen was pre-
served in alcohol. Dorsal body blackish brown, ventral
surface light khaki. Dorsal head and neck coincide with
the dorsal body, with a pair of light spots on each side of
the neck and on the back of head; dorsum without distinct
blotches; venter immaculate, without any dark stripes or
scattered spots; ventral surface of tail with a dark longi-
tudinal stripe and blotches; ventral scales with dark out-
ermost corners.

Variations for population from Mt. Gongga, Lud-
ing County. Measurements of other specimens are given
in Table 4. Male with relatively longer tail (TaL/TL are
distinctly larger in the adult male). Adult female GXNU
DLR194 displays many scattered brown spots on the ven-
ter, and with distinct light-yellow blotches on the left and
right sides of the neck, but in adult male GXNU DLR195
the venter is immaculate without any dark spots. The
population of Mt. Gongga, Luding County differs slightly
from the type specimen in dorsal and ventral coloration
(dorsal body blackish brown vs brown, ventral surface
light khaki vs white), and differs from the lectotype ZISP
8823 in color spots at the base of the tail (absent vs pres-
ent) (Figs 3, 4, 5).

zse.pensoft.net

904 Liang, Y.-T. et al.: Calamaria berezowskii Gunther, 1896

Table 4. Main morphological characters of Calamaria berezowskii. Abbreviations are listed in the Materials and Methods. “—” indi-
cates missing data. Data of lectotype and syntype of C. berezowskii by Giinther 1896 and Nikolai Orlov.

Voucher No. ZISP 8823 - GXNU DLR194 GXNU DLR195 GXNU 20221215002
Type of specimen Lectotype Syntype - - -
Sex 3 - 3 2
Ontogenetic Adult Adult Adult Adult Juvenile
Preocular 1/1 1 1/1 1/1 -
Postocular 1/1 1 1/1 1/1 -
Supralabials 4/4 4 4/4 4/4 4/4
Infralabials 5/5 - 5/5 5/5 5/5
Dorsals 13-13-13 - 13-13-13 13-13-13 13-13-13
Ventrals 155 167 2+149 2+165 153
Subcaudals 22 14 25 16 16
Tailspot 2+1 - Absent Absent Absent
TL (mm) 290 245 248 305 123
SVL (mm) 271 220 222 288 115
TaL (mm) 19 25 26 17 8
TaL/TL 0.066 0.102 0.105 0.056 0.065
HL (mm) 8.0 7.0 8.1 8.3 -
HW (mm) 5.0 - 4.1 3.9 -
HH (mm) 4.0 - 3.5 4.0 -
HW/HL 0.625 - 0.506 0.470 -
HH/HL 0.500 - 0.432 0.482 -
EyeD (mm) 0.7 - 0.6 0.7 -
Eye-MouthD (mm) 1.0 - 0.9 1.1 -
A Cc

Figure 3. Calamaria berezowskii (GNXU DLR195): A. Dorsal head; B. Ventral head; C. Right view of head; D. Left view of head;
E. Dorsal middle body; F. Lateral middle body; G. Dorsal tail; H. Ventral tail; I. Dorsal view; J. Ventral view.

Detailed morphological comparisons. Detailed mor-
phological comparisons between Calamaria berezowskii
and 20 congeners of the genus from China and mainland
Southeast Asia are:

zse.pensoft.net

Calamaria berezowskii is distinguished from C. al-

biventer, C. lumbricoidea, C. prakkei, C. schlegeli, and
C. concolor by having fewer supralabials (4 vs 5—6), and
the 2™ and 3" supralabials shields touching the orbit (vs.

Zoosyst. Evol. 100 (3) 2024, 897-911

905

Figure 4. Calamaria berezowskii (GXNU DLR194) in life.

3 and 4" supralabials touching orbit), mental not touch-
ing anterior chin shields (vs. touching in C. albiventer,
C. lumbricoidea, C. prakkei and C. concolor).

Calamaria berezowskii 1s distinguished with C. Jovii,
C. nebulosa, C. thanhi and C. yunnanensis by having
presence of preocular scale (vs. absence of preocular
scale), eye diameter less than distance from eye to mouth
edge (vs. reverse condition in C. thanhi and C. nebulo-
sa), fewer ventral scales in males (149-155 vs 161—202
in C. lovii, 149-155 vs 184 in C. thanhi and 149-155 vs
167-184 in C. yunnanensis), more subcaudal scale in
males (22—25 vs 14—-20 in C. lovii, 22—25 vs 15—20 in
C. yunnanensis).

Calamaria berezowskii differs from C. sangi, C. gi-
alaiensis and C. buchi by having mental not touching
anterior chin shields (vs. touching), dorsal scales re-
duced to six rows at tail (vs. reduced to four rows),
fewer ventral scales (149-167 vs 190 in C. sangi, 149-
167 vs 191 in C. gialaiensis, 149-167 vs 221-236 in
C. buchi).

Calamaria berezowskii is distinct from C. arcana
and C. jinggangensis by having the eye diameter less
than the distance from the eye to mouth edge (vs. re-
verse condition in C. arcana and C. jinggangensis),
fewer ventral scales (149-167 vs 170-192 in C. arcana
and C. jinggangensis), more subcaudals in females (16
vs 12-14 in C. arcana and C. jinggangensis), a distinct-

ly different coloration (dark orange or orangish-red in
fresh specimens in C. arcana and C. jinggangensis),
presence of dark outermost corners on ventral scales
(vs. absence in C. arcana and C. jinggangensis), a
faint dark line along each side of the back present (vs.
opposite condition in C. arcana and C. jinggangen-
sis), a dark longitudinal line or scattered spots on the
underside of tail present (vs. absent in C. arcana and
C. jinggangensis).

Calamaria berezowskii differs from C. abramovi by
having the rostral wider than high (vs. width equal to high
in C. abramovi), fewer ventral scales in males (149-155
vs 159 in C. abramovi), dorsal scales reduced to six rows
at tail (vs. reduced to four rows in C. abramovi), tail
ends in obtuse point (vs. sharp point in the end of tail in
C. abramovi), and a distinctly different coloration (body
black with yellow-orange spots on venter in C. abramovi).

Calamaria berezowskii differs from C. andersoni by
having eye diameter smaller than distance from eye to
mouth edge (vs. reverse condition in C. andersoni), fewer
ventral scales in males (149-155 vs 171 in C. andersoni),
dorsal scales reduced to six rows at tail (vs. reduced to
four rows in C. andersoni), light blotches on neck present
(vs. absent in C. andersoni).

Calamaria berezowskii differs from C. septentriona-
lis by having eye diameter less than distance from eye to
mouth edge (vs. reverse condition in C. septentrionalis),

zse.pensoft.net

906 Liang, Y.-T. et al.: Calamaria berezowskii Gunther, 1896

f

a

«

= ae

——

Se +

&

Figure 5. Lectotype ZISP 8823 of Calamaria berezowskii: A. Right view of head; B. Left view of head; C. Dorsal view; D. Ventral view.

zse.pensoft.net

Zoosyst. Evol. 100 (3) 2024, 897-911 907

Mas. Zool Ac. Phe. 1606.

eet

Calamaria berezowskii, sp. nu.
(PL, 1 fig. A.)

Rostral shield reverted to, and distinctly visible on, the
upper side ofthe head. Vertical as broad as long, about four
times as wide as the supraocular; one ante-, and one post-
ocular. Four upper labials; the first pair of lower labials meeting
behind the symphysial. Two pairs of chin-shields, without a
central shield between them, the anterior not much longer
than the posterior. Orbit narrower than its distance from the
labial margin. Ventrals 155, 167; subcaudals 22, 14. Tail rather
obtuse, with a conical end. Upper parts brown, with a faint
dark line along each side of the back, and another running
along the meeting edges of the two outer series of scales.
Lower parts, and the lower half of the outer-most row of scales
uniform white. A more or less distinct yellowish collar, about
five scales distant from the head. Two pairs of small white
spots may be present or absent on the tail.

Wotal length . 2... 290 mill. 245 mill.
Length of head... $8 , ce oy,
i Apc le as ve ane 20

Two specimens from Lun-ngan-fu (Berzzowsk1, 1893).

This species is allied to its compatriot, Calamaria septentrio-
nalis, but has an entirely different rostral shield. In this respect
it comes nearer to Calamaria siamensis, which has a narrower
vertical shield, and variegated abdomen.

Figure 6. Original description of Calamaria berezowskii Gunther, 1896: A. Drawing; B. Description.

zse.pensoft.net

908

mental separated from anterior chin shields (vs. reverse
condition in C. septentrionalis), not gradually tapering at
tip (vs. tail tip broadly rounded in C. septentrionalis), high-
er number of subcaudals (22—25 vs 15—19 in males and 16
vs 6-11 in females), absence of black stripe on venter of
tail (vs. venter of tail with broad and distinct median black
stripe in C. septentrionalis).

Calamaria berezowskii differs from C. pavimentata
by having the rostral shield width larger than high (vs.
reverse condition in C. pavimentata), the eye diameter
less than the distance from eye to mouth edge (vs. reverse
condition in C. pavimentata), tail not tapering anteriorly
and abruptly pointed at tip (vs. tail tapering gradually to a
point in C. pavimentata), dorsal scales reduced to six rows
at tail (vs. reduced to four rows in C. pavimentata), dor-
sum of body with two faint dark lines (vs. distinct narrow
and dark longitudinal lines on dorsum in C. pavimentata).

Calamaria berezowskii differs from C. dominici by
having eye diameter less than eye-mouth distance (vs.
opposite situation in C. dominici), mental separated from
anterior chin shields (vs. mental touching tip of right an-
terior chin shield in C. dominici), dorsal scales reduced
to six rows at tail (vs. reduced to four rows in C. domi-
nici), fewer ventral scales in females (153-165 vs 174 in
C. dominici), absence of blotches on dorsum (vs. present
irregular yellow blotches on dorsum in C. dominici), ven-
ter immaculate without dark stripes (vs. ventral side dark
with yellow blotches and bands).

Calamaria berezowskii differs from C. strigiventris by
having posterior chin shields meeting in midline (vs. di-
verging or meeting only in anteriorly in C. strigiventris),
dorsal scales reduced to six rows at tail (vs. reduced to
four rows in C. strigiventris), lower number of subcaudals
(22—25 vs 29-33 in males and 16 vs 20-30 in females in
C. strigiventris), venter immaculate (vs. presence of three
interrupted longitudinal black stripes in C. strigiventris),
tail not flattened, tapering and bluntly pointed at tip (vs.
slowly tapering anteriorly, then abruptly tapering to a
point in C. strigiventris).

Diagnosis. Calamaria berezowskii can be distin-
guished from all other congeneric species by having the
following combination of morphological characters: 1)
rostral shield width larger than high; 2) prefrontal shorter
than frontal, touching the first and second supralabials;
3) frontal hexagonal, length longer than width; 4) mental
not touching anterior chin shields; 5) two pairs of chin-
shields, the anterior not much longer than the posterior,
less than 1/2 of the posterior chin shield meets in the
midline; 6) eye diameter less than eye-mouth distance;
7) single preocular, single postocular; 8) four supralabi-
als, second and third supralabials entering orbit; 9) five
infralabials, first three touching anterior chin shields, first
pair of supralabials touching each other; 10) six scales
and shields surrounding the paraparietals; 11) dorsal
scales smooth, DSR 13-13-13 (n = 4); 12) dorsal scales
reduced to six rows above last subcaudal at tail; 13) 149
(+2 preventrals)}-155 ventrals in the males (n = 2), 153-
165 (+2 preventrals) in the females (n = 2); 14) 22—25

zse.pensoft.net

Liang, Y.-T. et al.: Calamaria berezowskii Gunther, 1896

subcaudals in the males (n= 2), 16 in the females (n = 2),
all paired; 15) anal plate single and intact; 16) dorsum of
body and tail blackish-brown or brown, with a faint dark
line along each side of the back; 17) a more or less dis-
tinct yellowish collar; 18) 2—3 pairs of small white spots
may be present or absent on the tail; 19) ventral scales of
body light khaki or white, with a dark longitudinal line or
scattered spots on the underside of tail; 20) two outermost
dorsal scale rows light khaki with upper margins partly
dark pigmented; 21) tail relatively short (5.6-10.5% of
the total length), not flattened, slowly tapering to obtuse,
with a conical end.

Etymology. Named after collector of type specimen,
Russian traveler and zoologist. Michael Berezowski. For
common name, we suggest “)!| PEPYS” (Chinese),
“Berezowski’s Reed Snake” (English).

Distribution and ecology. Terra typica of Calamaria
berezowskii is Lun-ngan-fu (now Long’an Town of Ping-
wu County) (Gunther 1896). It is known from western S1-
chuan Province, China, including Pingwu County, Shim-
ian County, Baoxing County, Luding County, Mt. Emei
and Pingshan County (Zhao et al. 1998; Zhao 2003; this
study). These localities are all on the eastern edge of the
Qinghai-Tibet Plateau bordering Sichuan Basin. In Lud-
ing County, it was found at an elevation of 1680-1827
meters, surrounded by mountainous evergreen broad-
leaved forest belt and evergreen broad-leaved deciduous
broad-leaved mixed forest belt (Fig. 7). It has a typical
subtropical-based vertical natural belt spectrum, with big
altitude differences, and abundant rare plant and animal
resources (Wang et al. 2023).

Discussion

Morphologically, specimens of C. berezowskii col-
lected in Luding County, Sichuan Province are a close
match to the type specimens collected in Pingwu Coun-
ty, Mianyang City of Sichuan Province (Gunther 1896).
For example, there was no significant difference in the
number of ventral and subcaudal scales; presence of a
more or less distinct yellowish collar; one preocular and
one postocular; presence of faint dark line along each
side of the dorsum. Geographically, Luding County and
Pingwu County are both located in the western region
of the Sichuan Province, which lies on the eastern slope
of the Qinghai-Tibet Plateau bordering the Sichuan Ba-
sin. Even though the samples of Calamaria used by this
study do not originate from the type locality, the mor-
phology of the voucher specimens matches the original
description by Gunther (1896), so we confirm that the
Calamaria specimens collected in Luding County, Sich-
uan Province are C. berezowskii and recover the validity
of C. berezowskii.

Due to their subterranean mode of life, digging be-
havior and mysterious habits, Calamaria species are not
often encountered in the wild. However, in the last five
years, five species of Calamaria were described, which

Zoosyst. Evol. 100 (3) 2024, 897-911

909

Figure 7. Macrohabitat of Calamaria berezowskii in Luding County, Sichuan Province, China.

once more shows that the diversity of the genus Cala-
maria is still highly underestimated. In addition, the
formal redescription of C. berezowskii brings the total
number of species of the genus Calamaria in China to
seven, namely C. pavimentata, C. septentrionalis, C. yun-
nanensis, C. andersoni, C. arcana, C. jinggangensis, and
C. berezowskii (Cai et al. 2023; Uetz et al. 2023). At the
same time, finding additional specimens of C. berezowskii
from the type locality and surrounding areas would allow
for a greater understanding of its intraspecific variation.
C. berezowskii was found at an elevation of 1680—
1827 meters, which is higher than the altitude of all
previously known Calamaria in China (vs.175—1520
m a.s.l.). The discovery of this new species increases
the elevation of all known Calamaria species in China
and suggests that the actual habitat of Calamaria spe-
cles may span a greater range of elevations. The Heng-

duan Mountains in southwest China are a biodiversity
hot spot, harboring a high percentage of endemic biota
(Hu et al. 2012; Wu et al. 2013; Sun et al. 2017; Qiao et
al. 2022). For snakes, the high altitude, steep terrain and
varying climatic conditions of the Hengduan Mountains
are important factors limiting their distribution and are
conducive to species migration and isolation, accelerat-
ing gene flow and promoting species adaptation to the
environment and species formation. Therefore, studying
this species is beneficial for understanding the influence
of geographical factors and the ecological environment
on species formation and evolution.

We update the key to Calamaria in China, which is
based on Zhao (2006), Yeung et al. (2022) and Cai et al.
(2023). The identification of this key is only for refer-
ence. Species should be identified by comparing more
morphological characteristics or molecular sequences.

Key to the identification of the genus Calamaria from China

1 FRE OCU aA SEN sense) vate ae erneme ee epee ear e ware aenry ecm ery ees pase xa Dena nt Pe nay died ee UREN NPN ORR ream nese xe Rare er Calamaria yunnanensis
— Pee OC Ml evarO le Sein Seeesuisure ste: Mee syns olubtuh 2 ees oldies oe le Masa Ue een eo Bae lai ee Lee meee in mal, ces me ALLL Sze 2
2 Dersal=seales reclice datote ur mowsionatallWatuleaStSUDCall Cal S xen uta ans a eens re tte bree le io Viel os bn 20 Ae S
- Dorsal:sealesaregucecsto more: thantouterowsson tallral: |aStsSubCallGlalS cer war se yi di, PINAR Naina y/ MR NR eT 5
S) Pie hitniness/ BlOtches=Ongneck ran celal a SOit: sad earct Ocul nee eared eens ate ed ON Peerage Reese ee Calamaria andersoni
- PreSenGevon IhShitchiMSS7/iDIOLCNES OM ME CKROR all fics cas acicnatieauewrtndeogs tans acts sebravesdiaeeciesslenesleemiginh sills aucdeaa tibia dclinaget sau dude acacace doasehe 4
A Mail tapers: SCAG Al Vy ROPAP OV 00s eee PE EP he, 0... ch ROPER eer. Sree SEPT ORE the J. chee SON ERERE er Calamaria pavimentata

Tail not tapering, broadly rounded on tip...................0006

Seed ect ce god cabs aera ane eeiady aU nae eae Calamaria septentrionalis

zse.pensoft.net

910

Liang, Y.-T. et al.: Calamaria berezowskii Gunther, 1896

5 Most of the posterior chin shield meets in the midline, absence of collar or spots in nuchal region ...............0::eeeeeeeee eee

chit, eI SE ee RE en Calamaria jinggangensis

- Less than 1/2 of the posterior chin shield meets in the midline, present indistinct light ring or one pair of spots in nu-

CAMO OUOU Te she. ioe EMER An: Be ete SAA Ta ts RA eRe uee ARR he e MRE ce pec dee eae AMEN tee 6
6 Ventral scales immaculate, without dark outermost corners and pigmentation anteriorly.............::.0 Calamaria arcana
- Two outermost dorsal scale rows light khaki, with upper margins partly dark pigmented.............. Calamaria berezowskil
Acknowledgements Inger RF, Marx H (1965) The systematics and evolution of the oriental col-

This study was supported by Science and Technology
Programme Project of Guangxi, China (AD21220058),
the National Natural Science Foundation of China (NSFC
32200379), the Natural Science Foundation of Guangx1,
China (2023GXNSFBA026309), and Survey and Assess-
ment of Priority Areas for Biodiversity Conservation in the
Terrestrial Area of Guangxi, China (2022-2023). It is part-
ly supported by the State Themes of ZISP 122031100282-
2 and grant no.075-15-2021-1069 by the Ministry of Sci-
ence and Higher Education of the Russian Federation.

We thank Xu Zhang and Congcong Du for providing
specimens, and thanks to Konstantin Milto for the help
with photographs of type specimens. We would also like
to thank Xin Wang and the reviewers for their valuable
comments on the manuscript.

References

Alencar LRV, Quental TB, Grazziotin FG, Alfaro ML, Martins M,
Venzon M, Zaher H (2016) Diversification in vipers: Phylogenet-
ic relationships, time of divergence and shifts in speciation rates.
Molecular Phylogenetics and Evolution 105: 50-62. https://doi.
org/10.1016/j.ympev.2016.07.029

Burbrink FT, Lawson R, Slowinski JB (2000) Mitochondrial DNA
phylogeography of the polytypic North American rat snake (E/aphe
obsoleta): A critique of the subspecies concept. Evolution 54(6):
2107-2118. https://doi.org/10.1111/j.0014-3820.2000.tb01253.x

Cai B, Jiang JP, Wu YY, Huang S, Fei DB, Ding L (2023) A new spe-
cies of Calamaria (Reptilia: Serpentes: Colubridae) from Western
Jiangxi Province, China. Russian Journal of Herpetology 30(2):
101-111. https://doi.org/10.30906/1026-2296-2023-30-2-101-111

Dowling HG (1951) A proposed standard system of counting ventrals in
snakes. British Journal of Herpetology 1: 97-99.

Grazziotin FG, Zaher H, Murphy RW, Scrocchi G, Benavides MA,
Zhang YP, Bonatto SL (2012) Molecular phylogeny of the new
world Dipsadidae (Serpentes: Colubroidea): a reappraisal. Cladistics
28(5): 437-459. https://doi.org/10.1111/j.1096-0031.2012.00393.x

Grismer LL, Kaiser H, Yaakob NS (2004) A new species of reed snake
of the genus Calamaria H. Boie, 1827, from Pulau Tioman, Pahang,
West Malaysia. Hamadryad 28: 1-6.

Gunther A (1896) Report on the collections of reptiles, batrachians and
fishes made by Messrs Potanin and Berezowski in the Chinese prov-
inces Kansu and Sze-chuen. Ezhegodnik. Zoologicheskogo Muzeya
Akademii Nauk SSSR 1896: 199-219.

Hu JH, Li C, Xie F, Jiang JP (2012) Endemic amphibians and their dis-
tribution in China. Asian Herpetological Research 3(2): 163-171.
https://doi.org/10.3724/SP.J.1245.2012.00163

zse.pensoft.net

ubrid snakes of the genus Calamaria. Fieldiana. Zoology 49: 1-304.

Kumar S, Stecher G, Li M, Knyaz C, Tamura K (2018) MEGA
X: Molecular evolutionary genetics analysis across computing
platforms. Molecular Biology and Evolution 35(6): 1547-1549.
https://doi.org/10.1093/molbev/msy096

Lanfear R, Calcott B, Ho SYW, Guindon S (2012) PartitionFinder:
Combined selection of partitioning schemes and substitution models
for phylogenetic analyses. Molecular Biology and Evolution 29(6):
1695-1701. https://doi.org/10.1093/molbev/mss020

Lanfear R, Frandsen PB, Wright AM, Senfeld T, Calcott B (2017)
PartitionFinder 2: New methods for selecting partitioned models
of evolution for molecular and morphological phylogenetic analy-
ses. Molecular Biology and Evolution 34(3): 772-773. https://do1.
org/10.1093/molbev/msw260

Lawson R, Slowinski JB, Crother BI, Burbrink FT (2005) Phylogeny
of the Colubroidea (Serpentes): New evidence from mitochondrial
and nuclear genes. Molecular Phylogenetics and Evolution 37(2):
581-601. https://doi.org/10.1016/j.ympev.2005.07.016

Lee JL (2021) Description of a new species of Southeast Asian reed
snake from northern Laos (Squamata: Colubridae: Genus Cala-
maria F. Boie, 1827) with a revised diagnosis of Calamaria yunnan-
ensis Chernov, 1962. Journal of Natural History 55(9-10): 531-560.
https://doi.org/10.1080/00222933.2021.1909165

Nagy ZT, Lawson R, Joger U, Wink M (2004) Molecular systematics
of racers, whipsnakes and relatives (Reptilia: Colubridae) using mi-
tochondrial and nuclear markers. Journal of Zoological Systematics
and Evolutionary Research 42(3): 223-233. https://doi.org/10.1111/
j.1439-0469.2004.00249.x

Nguyen TQ, Koch A, Ziegler T (2009) A new species of reed snake,
Calamaria Boie, 1827 (Squamata: Colubridae), from central Viet-
nam. Hamadryad 34(1): 1-8.

Orlov NL (2009) A new species of the genus Calamaria (Squama-
ta: Ophidia: Colubridae) from the Central Highlands (Ngoc Linh
Nature Reserve, Ngoc Linh Mountain, Kon Tum Province), Viet-
nam. Russian Journal of Herpetology 16(2): 146-154. https://doi.
org/10.30906/1026-2296-2009-16-2-146-154

Orlov NL, Nguyen QT, Nguyen TT, Ananjeva NB, Ho TC (2010) A
new species of the genus Calamaria (Squamata: Ophidia: Colubri-
dae) from Thua Thien-Hue Province, Vietnam. Russian Journal of
Herpetology 17(3): 236-242. https://doi.org/10.30906/1026-2296-
2010-17-3-236-242

Poyarkov NA, Van Nguyen T, Orlov NL, Vogel G (2019) A new spe-
cies of the genus Calamaria Boie, 1827 from the highlands of
the Langbian Plateau, Southern Vietnam (Squamata: Colubri-
dae). Russian Journal of Herpetology 26(6): 335-348. https://do1.
org/10.30906/1026-2296-20 1 9-26-6-335-348

Qiao J, Jia GQ, Zhou HM, Gong L, Jiang Y, Xiao NW, Gao XQ,
Wen AX, Wang J (2022) Mammal and bird diversity recorded
with camera traps in Gongga Mountain National Nature Reserve,

Zoosyst. Evol. 100 (3) 2024, 897-911

Sichuan, China. Shengwu Duoyangxing 30(2): 20395. https://doi.
org/10.17520/biods.2020395

Quah RV, Tan YH, Mubarak NM, Khalid M, Abdullah EC, Nolas-
co-Hipolito C (2019) An overview of biodiesel production using re-
cyclable biomass and non-biomass derived magnetic catalysts. Jour-
nal of Environmental Chemical Engineering 7(4): 103219. https://
doi.org/10.1016/j.jece.2019.103219

Ronquist F, Teslenko M, Van Der Mark P, Ayres DL, Darling A, Hohna
S, Larget B, Liu L, Suchard MA, Huelsenbeck JP (2012) MrBayes
3.2: Efficient Bayesian phylogenetic inference and model choice
across a large model space. Systematic Biology 61(3): 539-542.
https://doi.org/10.1093/sysbio/sys029

Stamatakis A (2014) RaxML version 8: A tool for phylogenetic anal-
ysis and post-analysis of large phylogenies. Bioinformatics 30(9):
1312-1313. https://doi.org/10.1093/bioinformatics/btu033

Stuart BL, Heatwole H (2008) Country records of snakes from Laos.
Hamadryad 33(1): 97-106.

Sun H, Zhang JW, Deng T, Boufford DE (2017) Origins and evolution
of plant diversity in the Hengduan Mountains, China. Plant Diversi-
ty 39(4): 161-166. https://doi.org/10.1016/j.pld.2017.09.004

Uetz P, Freed P, HoSek J (2023) The Reptile Database. http://www. rep-
tile-database.org [Accessed on 9 May 2023]

Wallach V, Williams KL, Boundy J (2014) Snakes of the World: A Cat-
alogue of Living and Extinct Species. CRC press, 1* edn., 1237.
https://doi.org/10.1201/b16901

Wang XM, Li Y, Liao R, Liu YX, Zhang XA, Qiao J, Wang Y, Wang X,
Liu SY (2023) Species diversity of small mammals on the north and
south slopes of Gongga Mountain National Nature Reserve. Sichuan
Linye Keji 44(6): 1-8. https://doi.org/10.12172/202307240001

Weinell J, Leviton AE, Brown RM (2021) A new species of reed snake,
genus Calamaria (Colubridae: Calamariinae), from Mindoro Island,
Philippines. Philippine Journal of Systematic Biology 14(2): 1-14.
https://doi.org/10.26757/pjsb2020b14006

Wu YJ, Colwell RK, Rahbek C, Zhang CL, Quan Q, Wang CK, Lei FM
(2013) Explaining the species richness of birds along a subtropical
elevational gradient in the Hengduan Mountains. Journal of Bioge-
ography 40(12): 2310-2323. https://doi.org/10.1111/jbi. 12177

Yang DT, Rao DQ (2008) Amphibia and Reptilia of Yunnan. Kunming:
Yunnan Science and Technology Press.

Appendix 1
Examined specimens (n = 6)

Calamaria berezowskii (n = 4). China: Sze-chuen (now
Sichuan Province): Lun-ngan-fu (Long’an Fu, now
Long’an Town of Pingwu County): ZISP 8823 (lec-
totype, adult male); Sichuan: Mt. Gongga, Moxi
Town, Luding County, Ganzi Tibetan Autonomous

a

Yang JH, Zheng X (2018) A new species of the genus Calamaria (Squa-
mata: Colubridae) from Yunnan Province, China. Copeia 106(3):
485-491. https://doi.org/10.1643/CH- 17-663

Yeung HY, Lau MWN, Yang JH (2022) A new species of Calamaria
(Squamata: Colubridae) from Guangdong Province, southern Chi-
na. Vertebrate Zoology 72: 433-444. https://doi.org/10.3897/vz.72.
©84516

Zaher H, Grazziotin FG, Cadle JE, Murphy RW, Moura-Leite JCD,
Bonatto SL (2009) Molecular phylogeny of advanced snakes (Ser-
pentes, Caenophidia) with an emphasis on South American Xeno-
dontines: A revised classification and descriptions of new taxa. Pa-
péis Avulsos de Zoologia 49(11): 115-153. https://doi.org/10.1590/
S003 1-10492009001100001

Zhang H, Fei DB, Zhao C, Liu S, Cai B, Guo P, Wu YY (2023) Cala-
maria arcana found in Chenzhou, Hunan. Chinese Journal of Zoolo-
gy 58(4): 615-621. https://doi.org/10.13859/}.cjz.202304014

Zhao EM (2003) Coloured Atlas of Reptiles of Sichuan. China Forestry
Press: Beijing, China.

Zhao EM (2006) Snakes of China. Hefei, China: Anhui Science and
Technology Press.

Zhao EM, Huang MH, Zong Y (1998) Fauna Sinica Reptilia, Vol. 3:
Squamata: Serpentes. Beijing. Science Press 1998(522): 1-8.

Ziegler T, Quyet LK (2005) A new species of reed snake, Calamaria
(Squamata: Colubridae), from the Central Truong Son (Annamite
Mountain range), Vietnam. Zootaxa 1042(1): 27-38. https://doi.
org/10.11646/zootaxa. 1042.1.2

Ziegler T, Hendrix R, Vu NT, Vogt M, Forster B, Dang NK (2007) The
diversity of a snake community in a karst forest ecosystem in the
central Truong Son, Vietnam, with an identification key. Zootaxa
1493(1): 1-40. https://doi.org/10.11646/zootaxa.1493.1.1

Ziegler T, Nguyen VS, Nguyen QT (2008) A new reed snake of the
genus Calamaria Boie (Squamata: Colubridae) from Vietnam. Cur-
rent Herpetology 27(2): 71-80. https://doi.org/10.3105/1881-1019-
272.71

Ziegler T, Tran VA, Babb RD, Jones TR, Moler PE, Van Devender
RW, Nguyen TQ (2019) A new species of reed snake, Calamaria
Boie, 1827 from the Central Highlands of Vietnam (Squamata:
Colubridae). Revue Suisse de Zoologie 126(1): 17—26. https://doi.
org/10.5281/zenodo.2619512

Prefecture: GXNU DLR195 (adult female); GXNU
DLR194 (adult male); GXNU 20221215002 (juve-
nile female).

Calamaria jinggangensis (n = 2). China: Guangxi:
Quanzhou County, Guilin) GXNU_ 20210909007
(adult male); Longsheng County, Guilin: GXNU
20220613012 (adult male).

zse.pensoft.net