Journal of Threatened Taxa |
www.threatenedtaxa.org | 26 June 2023 | 15(6): 23315–23326
ISSN 0974-7907
(Online) | ISSN 0974-7893 (Print)
https://doi.org/10.11609/jott.8486.15.6.23315-23326
#8486 | Received 19
April 2023 | Final received 12 May 2023 | Finally accepted 11 June 2023
Addition of four new records of
pit vipers (Squamata: Crotalinae) to Manipur, India
Premjit Singh Elangbam
1, Lal Biakzuala 2, Parag Shinde
3, Ht. Decemson 4, Mathipi Vabeiryureilai 5
& Hmar Tlawmte
Lalremsanga 6
1 Wildlife Explorers, Toubul, Bishnupur, Manipur
795126, India.
2,4,5,6 Developmental Biology and
Herpetology Laboratory, Department of Zoology, Mizoram University, Aizawl,
Mizoram 796004, India.
3 Asiyatic
Enclave, Malhar 203, Valtak
Nagar, Thane, Maharashtra 404606, India.
1 premjitdanao@gmail.com, 2
bzachawngthu123@gmail.com, 3 snakeparag5@gmail.com, 4
decemsonht@gmail.com, 5 m.vabeiryureilai@gmail.com, 6 htlrsa@yahoo.co.in
(corresponding author)
Editor: S.R. Ganesh, Kalinga
Centre for Rainforest Ecology, Shivamogga, India. Date of publication: 26 June 2023 (online & print)
Citation:
Elangbam, P.S., L. Biakzuala,
P. Shinde, Ht. Decemson, M. Vabeiryureilai
& H.T. Lalremsanga (2023). Addition of four new
records of pit vipers (Squamata: Crotalinae) to
Manipur, India. Journal of Threatened Taxa 15(6): 23315–23326. https://doi.org/10.11609/jott.8486.15.6.23315-23326
Copyright: © Elangbam
et al. 2023. Creative Commons Attribution 4.0 International License. JoTT allows
unrestricted use, reproduction, and distribution of this article in any medium
by providing adequate credit to the author(s) and the source of publication.
Funding: This work is funded by
the Science and Engineering Research Board (SERB) project (EEQ/2021/000243),
Department of Science and Technology, Government of India and MV would like to
thank a Research Promotion Grant received from Mizoram University (No. A.
49016/1/2019-Conf/PF; dt. 3rd February, 2023 and
49016/1/2019-Conf/PF; dt. 3rd March, 2023).
Competing interests: The authors declare no competing interests.
Author details: Premjit
Singh Elangbam is a BCom
graduate currently engaging in wildlife photography & research in Manipur.
He has contributed photographs and articles in many books on flora & fauna
of Manipur especially Indian birds. He has involved in many efforts in
conservation research conducted by Forest Dept., Govt. of Manipur. Now, he is
secretary of Wildlife Explorers Manipur, an organization fully devoted to
wildlife photography based on conservation & research in Manipur. Lal Biakzuala
is an emerging researcher in the field of herpetological taxonomy and
systematics, natural history, molecular phylogenetics, and conservation. He is
currently pursuing his PhD degree focussing on the
molecular systematics of elapid snakes, and is also currently working under
ICMR funded project for estimating the incidence, mortality, morbidity and
economic burden due to snakebites in India. Parag
Shinde is a commerce graduate and has also finished a photography
school. He is passionate about reptile photography, herpetological related
field investigations and animal rescuing. H.t.
Decemson is currently doing his Ph.D. work on
primates and also engaged as JRF under the DST-SERB project on the taxonomy,
ecology and distribution of amphibians and reptiles in Mizoram. He also
extended his work on the herpetofauna of Manipur State in collaboration with
the Department of Forest, Sanjenthong, Imphal, Government of Manipur. M. Mathipi Vabeiryureilai
is an assistant professor with an expertise in cancer biology,
evaluation of medicinal plants, taxonomy and molecular phylogeny of various
taxa. H.t. Lalremsanga
is a professor and a pioneer in the field of herpetology in Mizoram. Currently,
he is teaching post-graduate students, guiding research scholars, and
overseeing major projects funded by the Department of Science and Technology -
Science and Engineering Research Board (DST-SERB) and Department of
Biotechnology (DBT) under the Ministry of Science and Technology, the Defence Research and Development Organisation
(DRDO) under the Ministry of Defence, Indian Council
of Medical research (ICMR), The Habitats Trust (THT), Noida and the National
Mission for Himalayan Studies (NMHS), Government of India.
Author contributions: PSE, HD, PS did the field-based investigations and sampling; LB performed data analyses, and wrote the first draft of the manuscript with HTL; MV carried out laboratory and technical works;
HTL conceived the study, review and editing, and approval of the manuscript prior submission. All authors reviewed
and agreed upon the final
version.
Acknowledgements: We are grateful to the chief
wildlife warden of forest department, Government of Manipur for the herpetofaunal collection permit in Manipur State (permit
no. #3/22/2018-WL [VolII]). We acknowledge the
funding agency DST-SERB sanction no. EEQ/2021/000243 for financial support.
Expressed thanks to Mr. Ngamsongbou Newmai and Mr. Langamba Ayekpam for fieldwork and we also appreciate for the
laboratory assistance of Mr. F. Malsawmdawngliana.
Abstract: In spite of the potential
richness in biodiversity, Manipur is still one of the least explored areas in
northeastern India. In this work, we report on our finding of four pit viper
species, namely, Trimeresurus mayaae, T. salazar, T.
erythrurus, and Protobothrops
jerdonii in Manipur during our herpetological
surveys in 2022. We affirm the identity of these specimens based on the
combination of standard morphological characters and mitochondrial cytochrome b
gene. Our study not only reports four new records of pit vipers for the first
time in Manipur, but also extends their respective known geographical ranges.
This information calls for further dissemination especially to health care
sector, as it is on venomous snakes that could be of medical importance as
regards human snakebites.
Keywords: morphology, new state report,
northeastern India, phylogenetics, Protobothrops,
range extension, systematics, Trimeresurus.
INTRODUCTION
Pit vipers are a group of
front-fanged venomous snakes that comprise various genera of vipers having heat
sensitive pit between each eye and nostril. Asian pit vipers that are
distributed across southern and southeastern Asia are known to inhabit several
types of habitats like wet, humid and montane forests, and also found in a few
dry forest habitats (Gumprecht et al. 2004). Recent
phylogeographic studies have largely addressed the taxonomic status of certain
groups of pit vipers like Trimeresurus complex
(Malhotra & Thorpe 1997, 2004; David et al. 2011; Mallik et al. 2021) and Protobothrops (see Guo et al. 2009).
Geographical diversity as well as sexual dimorphism often led to inaccurate
taxonomic assignment among the closely similar pit viper species (Malhotra
& Thorpe 1997, 2004). Consequently, pit vipers are still regarded to
contain several taxonomically complex species in India. Recently, many such
species complexes were revised including those from the Craspedocephalus
malabaricus and C. macrolepis
complexes (Mallik et al. 2021), the T. medoensis
complex (Rathee et al. 2022), and the T. albolabris complex (Chandramouli
et al. 2020; Mirza et al. 2020; Vogel et al. 2022).
Manipur is a hilly Indian state that lies at
the extreme east of the northeastern region of India bordering Nagaland in the
north, Mizoram in the south-east, Assam in the west, and Myanmar towards the
east and south-east. In spite of the potential richness in biodiversity,
zoological exploration in Manipur is rather limited (Roonwal
1948; Singh 1995; Pawar & Birand
2001) and there exist only a handful of snake faunal reports from the area till
date (e.g., Mathew 2005; Sinate et al. 2021, 2022; Elangbam et al. 2022). Although Wallach et al. (2014)
listed Manipur as the potential range of T. erythrurus,
there is no confirmatory records from the area, and none of the aforementioned
literature listed pit vipers from the snake fauna of Manipur except Ovophis monticola (see
Whitaker & Captain 2008). In this paper, we provide notes on our new addition
of pit vipers from Manipur based on our field documentations during our
herpetological surveys in 2022.
MATERIALS AND METHODS
Sampling and morphology
We conducted our field surveys
after obtaining herpetological specimen collection permit from the Forest
Department, Government of Manipur (permit no. #3/22/2018-WL [Vol-II]).
Morphological characters of the newly collected specimens were taken. We followed
Dowling’s (1951) method for counting ventrals (Ve), and we excluded the terminal scute
in counting the subcaudals (Sc). All the measurements are given in millimeter,
and other acronym used for the morphological characters are: snout-vent length,
SVL; tail length, TaL; inter-orbital distance
(measured on cephalic scale which was counted on a straight line between the
middle of the supraoculars), IOD; supralabials,
SL; infralabials, IL; and the bilateral characters are given as left/right. We
euthanized the live specimens using MS-222 according to the standardized
protocol (Conroy et al. 2009), and the procedure was approved by the
Institutional Animal Ethics Committee (IAEC) (Permission No. MZU-IAEC/2018/12).
All the specimens collected during this survey were deposited in the
Departmental Museum of Zoology, Mizoram University, India (MZMU). For
molecular analyses, we dissected the liver tissue from the freshly euthanized
specimens and stored in 95% ethanol at -20°C refrigerator. Distribution map was
prepared using QGIS v3.16.2.
DNA extraction and amplification
We employed mitochondrial
cytochrome b (cytb) gene for molecular based
identification. For this purpose, genomic DNA was extracted from the liver
tissue using QIAamp DNA Mini Kit following the
standard protocol provided. We amplified the fragment of cytb
gene using a published primer pairs (Snk) (Dubey et
al. 2009) under the thermal conditions of 94˚C for 3 min for initial
denaturation; 35 cycles for denaturation at 94˚C for 30 sec, annealing at 49˚C
for 40 sec, and extension at 72˚C for 30 sec; with the final extension of 72˚C
for 5 sec, and cooling at 4°C for 15 min. The amplified products were purified
using ThermoFisher ExoSAP-
IT PCR product clean-up reagent and sequenced at Barcode BioSciences,
Bangalore, India using Sanger’s dideoxy method. The newly generated sequences
were submitted to NCBI GenBank database (Benson et al. 2018) and obtained the
accession numbers (OQ968475–77).
Molecular analyses
We assembled the dataset mainly
based on the published datasets of Mirza et al. (2023) for Trimeresurus
and Guo et al. (2009) for Protobothrops,
and we obtained the published nucleotide sequences from the NCBI GenBank
database (Benson et al. 2018). We used the default parameter settings of MUSCLE
(Edgar 2004) for performing nucleotide alignment in MEGA 11 (Tamura et al.
2021). The flanking gaps created by the samples with shorter nucleotide
sequences are treated as missing data. We estimated the uncorrected p-distance
using complete deletion for gaps/missing data in MEGA 11 (Tamura et al. 2021).
We partitioned the assembled datasets (1,088 bp in
length for Trimeresurus; 847 bp in length for Protobothrops)
by codon position. The best partitioning schemes and nucleotide evolutionary
models selected by PartitionFinder v2 (Lanfear et al. 2017) under the Bayesian Information
Criterion are HKY+I for cytb pos1, TRN+G for cytb pos2, HKY+G for cytb pos3 in
the dataset of Trimeresurus; HKY+I+G for cytb pos1 and pos3, and TIM+I+G for cytb
pos2 in the dataset of Protobothrops. By
implementing the selected models, we performed the partitioned Bayesian
inference (BI) phylogeny separately for the two datasets in MrBayes
v3.2.5 (Ronquist
et al. 2012). We conducted four independent runs with one cold and three
hot chains for 10 million generations and sampling every 5000 generations. We
discarded 25% of the trees as burn-in cut off after examining the trace plots
generated by the MCMC runs in Tracer v1.7 (Rambaut et
al. 2018).
RESULTS AND DISCUSSION
In this report, we documented a
total of four pit viper species from Manipur (Image 1), and we established
their identity based on the combination of mitochondrial cyt
b gene and their respective morphological characters, including key diagnostic
features from relevant published literature (e.g., Guo et al. 2009; Zambre et al. 2009; Yang et al. 2011; Rathee
et al. 2022; Mirza et al. 2020, 2023).
Trimeresurus mayaae
Rathee et al., 2022 (Image 2)
The recently discovered Maya’s
Pit Viper, T. mayaae is a species genetically
close to T. medoensis as well as
morphologically similar to T. gumprechti of
Indochina (Rathee et al. 2022 and references therein).
Not much is known on the biology of this cryptic species, and its
description is based on specimens originating from the Northeast Indian States
of Mizoram and Meghalaya. This species, in fact represents the third new
species of the genus from India, to be described in the past three years (Rathee et al. 2022).
On 17 July 2022, we collected
both male (MZMU2971; Image 2a) and female (MZMU2970; Image 2b) individuals of T.
mayaae from Sadu Chiru
area nearby a waterfall in Kangpokpi District,
Manipur (24.740°N, 93.747°E; 1,139 m). A female individual was found
resting on top of tree branch at 2211 h and a male individual was found
crawling on the ground about 6 m away from where a female was collected at 2237
h. The microhabitat of the collection site was predominantly covered by thorny shrubs
like Solanum incanum, and bamboos such as Melocanna bacifera and
Chimonobambusa callosa.
These specimens were morphologically examined in the field and concorded to
the descriptions and diagnostic keys of Trimeresurus
mayaae fide Rathee et
al. (2022). The two newly collected specimens are characterized by the absence
of red and white post-ocular stripe in both specimens; iris greenish around the
periphery and adjoined by a prominent rust color around the center in male,
while it is greenish with partially blended by a rust color in female;
snout-vent length (SVL) 430 mm in male, and 310 mm in female; tail length (TaL) 97 mm in male, and 61 mm in female; dorsal scale row
(DSR) 19:19:15 in male, 21:19:15 in female; Ve 154 in
male, 152 in female; Sc 58 in male, 53 in female; inter-orbital distance (IOD)
measured in scale are 7 in male, and 8 in female; supralabials
(SL) 9/9 (left/right) in male, 10/9 in female; infralabials (IL) 10/11 in male,
10/10 in female, and the first pair contact with each other. The new specimens
updated the key diagnostic features in the lower limit of Ve
in male i.e., 154 vs. 157–162 (Rathee et al. 2022)
and female i.e., 152 vs. 153 (Rathee et al. 2022),
and also in the lower limit of Sc in female i.e., 53 vs. 54–55 (Rathee et al. 2022). These specimens also represent the
first record of the species from the Manipur State and extend the known
distribution range by ca. 146 km aerial distance north-eastward from the type
locality in Champhai, Mizoram and ca. 209 km aerial
distance towards SE from the paratypes locality in Umroi,
Meghalaya (Rathee et al. 2022) (Image 1).
Despite the fact that we are
utilizing a single gene (cyt b) in this study, our
reconstructed BI phylogram of Trimeresurus species
considerably concorded to the tree topology, particularly on the clades of
the subgenus “Trimeresurus” and “Viridovipera” fide Mirza et al. (2023) where
they combined four mitochondrial genes (cyt b, 16S
rRNA, 12S rRNA, and ND4). Accordingly, our cyt b gene
tree depicted the new T. mayaae specimen
(MZMU2970; Accession No. OQ968476) clustering alongside the type series of the
species by a well-supported Bayesian posterior probability support (PP = 0.99),
and is nested with the holotype from Mizoram (PP=1.00) (Figure 1). Moreover,
the specimen (MZMU2970) is showing 0.0–0.7% intraspecific genetic distance with
respect to the type series, and the least genetic distance was seen with the
holotype (0.0%) from Mizoram, India (see Table S1).
Trimeresurus salazar
Mirza et al.,
2020 (Image 3)
The Salazar’s Pit Viper, T. salazar is one of the recently discovered Trimeresurus in the past three years. The species
has been confirmed so far from central-southern Nepal, Southern Bhutan (Vogel
et al. 2022), Northeast Indian States of Arunachal Pradesh (type locality),
Assam (Mirza et al. 2020), Meghalaya (Rathee et al.
2021), Sikkim, Nagaland, and the other Indian States of Madhya Pradesh,
Jharkhand, Chhattisgarh, Bihar and Northern West Bengal (Vogel et al. 2022).
The species is distinguished from the congeners by the combined evidence of
mitochondrial 16S and ND4 genes, external morphology, dentary and hemipenial morphology (Mirza et al. 2020).
On 17 October 2022 at ca. 0900 h,
we encountered and captured a male individual coiled among bushes of tomato Solanum
lycopersicum inside a plantation surrounded by Mangifera indica, Lantana camara,
Psidium guajava, and Melocanna
bacifera at the foothills of Langol
Hill Reserve Forest, near the villages of Lamdeng and
Khundi, Imphal West
District, Manipur (25.227262°N, 93.939305°E; 2,170 m). The new specimen is
collected ca. 253 km aerial distance southeastward from the type locality at Pakke Tiger Reserve, Arunachal Pradesh (Mirza et al. 2020)
(Image 1). The other nearest occurrence record of the species is most likely
from Hamren, Karbi Anglong District in Assam (CESS604) ca. 174 km aerial
distance north-westward from the new collection site. Previously, the locality
for CESS604 was mistakenly provided as “Aizawl, Mizoram, India” by Vogel et al.
(2022) while it was actually captured from the Assam-Meghalaya interstate
border road (25.829°N, 92.556°E, 449 m) (Lalnunhlua
pers. comm.), and this clarification warrants the removal of Mizoram as part of
the known range of T. salazar until there is
further confirmatory work. The snake remained very active while capturing and
under temporary captivity for morphological examination. Based on the
morphological characters, we identified the specimen as a male T. salazar (MZMU3027) in agreement with literature
mentioning diagnostic keys (Mirza et al. 2020; Rathee
et al. 2020; Vogel et al. 2022). We obtained the following morphological
attributes: SVL 475 mm; TaL 110 mm; DSR 21:19:15; Ve 165, Sc 65; IOD 12; SL 11/11, and the first SL fused
with nasal scale; IL 12/12, and the first pair in contact with each other.
Although the lateral stripe on head, one of the taxon’s male key features fide
Mirza et al. (2020) is not evident in our specimen, we considered the
particular character as a phenotypically plastic trait as seen in T. mayaae (Rathee et al. 2022)
and T. popeiorum (Mirza et al. 2023) rather
than sexually dimorphic trait, and also in consideration of the low sampling
size in the original description (three males and one female) (See Mirza et al.
2020). Even so, with respect to T. erythrurus,
the species that usually lacks the red postocular
stripe in males, the specimen (MZMU3027) is distinct in having 19 mid-dorsal
scale rows vs. 23 in T. erythrurus (Mirza et
al. 2020).
Our cyt
b based BI phylogram also depicted a subclade of salazar
+ septentrionalis +caudornatus
that was conceived by Mirza et al. (2023); and the new T. salazar specimen (MZMU3027; Accession No. OQ968477)
clustered with the conspecies from Meghalaya, India
(CESS331) and Nepal (AM A100) with a well-supported branch (PP = 1.00)
(Figure 1). We uncovered 0.2–0.5% intraspecific genetic distance among T. salazar samples, and our sample shows 0.2%
genetic distance with respect to both of them (see Table S1).
Trimeresurus erythrurus
(Cantor,
1839) (Image 4)
The Red-tailed Pit Viper, T. erythrurus was originally described from the “Delta Gangeticum” which is apparently from the Indian Sunderbans of West Bengal state in the present time fide Deuti et al. (2021). The species is widely distributed in
parts of Southern Asia including Bhutan (Wangyal
2014; Deuti et al. 2021 and references therein),
Nepal, Bangladesh, Myanmar (Mahony et al. 2009; Wallach et al. 2014; Chan et
al. 2022), and the Indian states of Andhra Pradesh (Kakinada), West Bengal (Deuti et al. 2021), and Northeast India viz. Arunachal
Pradesh, Assam, Mizoram, Nagaland, and Sikkim (Whitaker & Captain 2008;
Wallach et al. 2014); possibly in Meghalaya and Manipur (see Wallach et al.
2014).
On 23 July 2022, at ca. 0700 h,
we documented a live individual of T. erythrurus from
a forest trail at Bishnupur District, Manipur
(24.632°N, 93.746°E; 920 m; Image 1) ca. 500 km aerial distance northeast from
the type locality in Sunderbans (Deuti
et al. 2021). Seeing the bulged abdomen, it was evident that the snake had
recently fed, possibly upon small mammal or a bird (Image 4a). Morphologically,
we identified the snake as female T. erythrurus in
having golden-yellow colored eye; greenish dorsum; white lateral stripe on the
head and the ventrolateral body; total length of 595 mm; Ve
168; Sc 51; and 23 mid-dorsal scale which are in agreement with the published
diagnostic characters of the species (Chen et al. 2020; Mirza et al. 2020; Deuti et al. 2021). We subsequently released back the snake
into the wild after taking measurements and scalation data. On 4 November 2022,
at ca. 1100 h, we encountered additional uncollected road killed male
individual at Noney town, Noney
District, Manipur (24.851°N, 93.617°E; 510 m asl.)
ca. 28 km aerial distance northwest from the point of our observation of the
live individual. As the snake was badly damaged and large amount of the scales
were eaten off by ants, we could barely examine the snake but nevertheless,
obtained the key features like white lateral stripe on the head and ventrolateral
body, 23 mid-dorsal scale rows, and Ve 158 (Image
4b). Given that we could not obtain genetic data for this species in Manipur,
we advocate DNA studies on the Manipur population to further corroborate the
present report.
Protobothrops jerdonii
(Günther,
1875) (Image 5)
The widely distributed P. jerdonii is presently found in southwestern China,
northeastern India, Nepal, northern Myanmar as well as northern Vietnam (see
Guo et al. 2009). The high level of variation in their ecology as well as morphology,
particularly in the scale counts and coloration has led to a controversial
intraspecific taxonomic treatment within P. jerdonii
(Guo et al. 2009). This has led some authors to recognise
the species as monotypic without any subspecies, while certain other authors
treat the taxon to be polytypic (see Guo et al. 2009) with a total of five
species or subspecies proposed so far within the P. jerdonii
complex. They are, apart from the nominotypical form jerdonii
Günther, 1875; xanthomelas Günther, 1889; melli Vogt, 1922; meridionalis
Bourret, 1935; and bourreti
Klemmer, 1963. Among these, only jerdonii,
bourreti and xanthomelas
were considered as valid subspecies by several authors like Hoge &
Romano-Hoge (1983), Golay et al. (1993), Orlov et al. (2001), and Gumprecht
et al. (2004).
On 18 July 2022 at 1949 h, we
encountered the specimen (MZMU3011) at Chawangkining
village nearby Zaimeng Lake, Kangpokpi
District, Manipur (25.204°N, 93.940°E; 1,725 m). The snake was captured while
crawling across a forest track covered by herbs and thorny shrubs like Mikania
micrantha, Mussaenda
spp., Girardinia diversifolia,
and Rubus spp., and surrounded by
bamboos such as Melocanna bacifera and Chimonobambusa
callosa. The collection site of the new specimen
is situated ca. 235 km aerial distance towards east from the type locality in
Khasi Hill, Meghalaya fide Wallach et al. (2014) (Image 1). In comparing the
descriptions and diagnostic keys from literature (e.g., Zambre
et al. 2009; Yang et al. 2011), we identified the specimen as male P. jerdonii in having SVL 414 mm; TaL
77; Ve 171; Sc 66; SL 7/8; IF 11/11; IOD 8; DSR
21:21:16. In considering the subspecies recognized by Gumprecht
et al. (2004), the Ve of our specimen (171) falls
within the range of P. jerdonii jerdonii (160–170) fide Yang et al. (2011), the Sc of
our specimen (66) enter the Sc range of both P. jerdonii
xanthomelas (54–67) and P. jerdonii
bourreti (65–72) fide Yang et al. (2011).
Moreover, the basal body color of our specimen is yellowish brown, and a series
of rhomboidal or irregularly shaped reddish-brown patches with black margins
are present along the dorsal body. Based on the morphological data alone, we
cannot certainly assign the new specimen (MZMU3011) to any of the three nominal
subspecies following the recent most diagnostic keys fide Yang et al. (2011).
Thus, we for now refrain from making subspecies level taxonomic identification
for the specimen.
Our reconstructed cyt b gene tree also depicted paraphyly amongst P. jerdonii as conceived by Guo et al. (2009) where they
sequenced four mitochondrial genes (cyt b, 12S rRNA,
16S rRNA, and ND4). Our sample (MZMU3011; Accession No. OQ968475) nested alongside
the conspecific sequences from Northeast India (V14) with a high branch support
(PP = 1.00), and these two formed a subclade with the other samples from China
(GP87 and KIZ012737) with high support (PP = 1.00) (Figure 2). Our estimated
genetic distance also showed 0.0–3.3% intraspecific distance between ours and
the other P. jerdonii samples; the least
genetic distance (0.0%) is seen with the Northeast Indian sample while the
highest (3.3%) is seen with respect to the samples from Huili,
Sichuan, China (GP11–GP14) (see Table S2).
Conclusion
As per the current knowledge,
only O. monticola has been listed as present
in Manipur (Whitaker & Captain 2008). In this work, we report the
occurrence of four new records of pit vipers in Manipur for the first time, and
simultaneously provide new information on their respective geographical
distribution ranges. Considering the topology in our phylogenetic
reconstruction and our P. jerdonii sample
(MZMU3011) that disclosed the overlapping of the conventional taxonomic keys
for the subspecies fide Yang et al. (2011), we are convinced that our study
corroborated the taxonomic statement on the species by Guo et al. (2009) where
they considered P. jerdonii as a monotypic and
paraphyletic species. In the scenario of T. mayaae,
both ours and the published phylogenies (Rathee et
al. 2022; Mirza et al. 2023) depicting the nesting of T. mayaae
next to T. medoensis. Nonetheless, combining
the meristic data of the type series (Rathee et al.
2022) and the new specimens, T. mayaae is
morphologically distinct in terms of the Ve counts of
male i.e., 154–162 vs. 138–149 in T. medoensis,
and female i.e., 152–153 vs. 141–143 in T. medoensis;
also in the mid-dorsal scale row i.e., 19–21 in T. mayaae
vs. 17 in T. medoensis.
The present work, however,
provides evidence for the occurrence of four additional species of pit vipers
in Manipur for the first time, and simultaneously provides new information on
their respective geographical distribution ranges. Thus, currently, five
species of pit vipers including P. jerdoni, T.
salazar, T. mayaae,
T. erythrurus, and O. monticola
are known from Manipur (Whitaker & Captain 2004; this work). We emphasise that this new data on the occurrence of four more
species of pit vipers in Manipur may be disseminated well in health care
sectors. This is so because these venomous snakes may be of medical importance
in terms of potential snakebites on humans (e.g., Kanwar 2018; Rai et al. 2021;
Ravikar et al. 2023), more so in forest fringe areas.
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