Journal of Threatened Taxa | www.threatenedtaxa.org | 26 June 2024 | 16(6): 25385–25399

 

ISSN 0974-7907 (Online) | ISSN 0974-7893 (Print) 

https://doi.org/10.11609/jott.8935.16.6.25385-25399

#8935 | Received 31 January 2024 | Final received 29 May 2024 | Finally accepted 13 June 2024

 

 

First record of two species of venomous snakes Bungarus suzhenae and Ovophis zayuensis (Serpentes: Elapidae, Viperidae) from India

 

Jason Dominic Gerard 1, Bitupan Boruah 2, V. Deepak 3  & Abhijit Das 4

 

1,2,4 Wildlife Institute of India, Chandrabani, Dehradun, Uttarakhand 248001, India.

3 School of Natural and Environmental Sciences, Newcastle University, Newcastle upon Tyne NE1 7RU, UK.

3 Natural History Museum, London SW7 5BD, UK.

1 jasondg95@gmail.com, 2 bitupan.kaz@gmail.com, 3 veerappandeepak@gmail.com, 4 abhijit@wii.gov.in (corresponding author)

 

 

Editor: S.R. Ganesh, Kalinga Foundation, Agumbe, India.            Date of publication: 26 June 2024 (online & print)

 

Citation: Gerard, J.D., B. Boruah, V. Deepak & A. Das (2024). First record of two species of venomous snakes Bungarus suzhenae and Ovophis zayuensis (Serpentes: Elapidae, Viperidae) from India. Journal of Threatened Taxa 16(6): 25385–25399. https://doi.org/10.11609/jott.8935.16.6.25385-25399

  

Copyright: © Gerard et al. 2024. 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: 1. Science and Engineering Research Board (SERB), Govt. of India; 2. National Geographic Society.

 

Competing interests: The authors declare no competing interests.

 

Author details: Jason D. Gerard is a researcher with interest in taxonomy and ecology of reptiles. Bitupan Boruah is a PhD scholar whose interests are systematics and biogeography of lesser-known fauna such as amphibians and reptiles. V. Deepak is a researcher whose work predominantly focuses on the evolution and systematics of lizards, snakes and turtles in southern Asia. Abhijit Das is a scientist and faculty involved in teaching, training and research in Herpetology.

 

Author contributions: JDG and BB equally contributed to this work. BB, AD and JDG carried out field work for this study. JDG and BB examined the specimens. AD and BB conceptualized the study. BB and DV contributed to the phylogenetic analyses and data verification. All authors contributed equally to the writing and editing of this manuscript.

 

Acknowledgements: We are grateful to the state Forest Department of Nagaland and Arunachal Pradesh for permitting us to carry out the study (via CWL/GEN/81/580 dated 31 August 2012 and CWL/GEN/355/2021/3178 dated 28th September 2021, respectively). We thank Taluk Rime (DFO, Anini), Kashok Solung (RFO, Anini), and Basant Taye (Forester, Anini) for their support during the field study. We are thankful to SERB-DST (CRG/2018/000790) for financial support and the National Geographic Society for the award of the National Geographic Explorer Grant (NGS-74044R-20). We are thankful to the director and dean of the Wildlife Institute of India, Dehradun for their support. We thank Neelam Dutta, Naitik G. Patel, Santanu Dey, Saurabh Dutta, Rajiv N.V., Gaurav Rai Pariyal, Hachi Molo, Dochi Meli, and Bhaiti for their help during fieldwork. We thank A. Madhanraj, Surya Prasad Sharma, Surya Narayanan, and Kumudani Bala Gautam for their help. We thank Fred Pluthero and section editor, JoTT for their valuable comments in the earlier version of this manuscript.

 

 

Abstract: We report Bungarus suzhenae Chen, Shi, Vogel, Ding & Shi, 2021 and Ovophis zayuensis (Jiang, 1977) for the first time from India. Specimens of B. suzhenae and O. zayuensis were collected during our field surveys in north (Arunachal Pradesh) and south (Nagaland-Manipur border) of the river Brahmaputra. Species identity was supported by partial cytochrome b (cyt b), and 16s mitochondrial gene. We provide a detailed morphological description and a key to the two genera of this region. This report extends the westernmost distribution of B. suzhenae by ca. 300 km from Myanmar, and the southernmost range of O. zayuensis by 170 km from Tibet. Until now eight species of Bungarus and only one Ovophis species have been reported from India. Ovophis species are recently reported to be medically important venomous snakes whose venom properties have not been investigated in depth.

 

Keywords: Krait, northeastern India, pit viper, range extension, taxonomy.

 

 

INTRODUCTION

 

Northeastern India is part of the Indo-Burma biodiversity hotspot (Tripathi et al. 2016) and home to approximately 21 species of venomous snakes from the families Viperidae and Elapidae (Ahmed et al. 2009; Captain et al. 2019; Mirza et al. 2020; Rathee et al. 2022; Biakzuala et al. 2023 and 2024). Biodiversity explorations are continuing to reveal new information on snake diversity (Elangbam et al. 2023; Biakzuala et al. 2024). The Elapid genus Bungarus Daudin, 1803 consists of 18 species (Uetz et al. 2024), distributed from Iran and Pakistan eastwards to China and Indonesia (Chen et al. 2021). Eight species of Bungarus have been reported from India, namely, B. andamanensis Biswas & Sanyal, 1978, B. caeruleus Schneider, 1801, B. sindanus Boulenger, 1897, B. walli Wall, 1907, B. lividus Cantor, 1839, B. fasciatus Schneider, 1801, B. niger Wall, 1908, and B. bungaroides Cantor, 1839, of which the latter four are found in northeastern India (Ahmed et al. 2009; Das & Das 2017; Das 2018; Biakzuala et al. 2021). Recently, Chen et al. (2021) described B. suzhenae from China that is currently known from the type locality Yingjiang County, Yunnan Province, China, and the adjacent Kachin State of Myanmar. 

There are eight species in the viperid genus Ovophis (Burger, 1981), namely: O. makazayazaya (Takahashi, 1922); O. jenkinsi Qiu et al., 2024; O. malhotrae Zeng, Li, Liu, Wu, Hou, Zhao, Nguyen, Guo & Shi, 2023; O. monticola (Gunther, 1864); O. okinavensis (Boulenger, 1892); O. tonkinensis (Bourret, 1934); and O. zayuensis (Jiang, 1977) (Malhotra et al. 2011; Zeng et al. 2023; Qiu et al. 2024; Uetz et al. 2024). The southernmost record of O. zayuensis is from Zayu, Motuo, Xizang province, China. During a herpetological field survey in northeastern India in 2012, 2022, and 2023, we came across one individual of Bungarus and two individuals of Ovophis. They were collected and based on morphological evaluation we identified them as B. suzhenae and O. zayuensis. Additionally, we generated new DNA sequences and compared them with published sequences of these species, including their types (B. suzhanae CIB 116088 from Yingjiang County, Yunnan Province, China, and O. zayuensis CIB 013375 from Chayu Co., Xizang, China), based on which we report here the first country records for India.

 

 

MATERIALS AND METHODS

 

We carried out field surveys during October 2012 in the Nagaland-Manipur border, between June and July 2022–2023 in Arunachal Pradesh, northeastern India (Figure 1). Collected specimens were fixed in formalin, washed, stored in 70% ethanol, and housed in the repository at the Wildlife Institute of India, Dehradun (WII-ADR1241, WII-ADR3316 and WII-ADR3491). Before fixation, live snakes were photographed, and liver tissues were collected and stored in absolute ethanol at -20°C.

 

Molecular phylogenetic analysis

Genomic DNA was extracted from liver tissue samples using DNeasy Blood and Tissue Kit (Qiagen™, Germany). For the Bungarus specimen (WII-ADR1241), we amplified and sequenced fragments of one mitochondrial gene, cytochrome b (cyt b) using the primers L14910 (5’-GACCTGTGATMTGAAAACCAYCGTTGT-3’) and H16064 (5’-CTTTGGTTTACAAGAACAATGCTTTA) (Burbrink et al. 2000). Polymerase Chain Reaction (PCR) conditions followed was initial denaturation at 95°C for five minutes, followed by 35 cycles of denaturation at 95°C for 45 sec, annealing at 54°C for 45 sec, and extension at 72°C for 55 sec. The final extension was at 72°C for 10 min. For the Ovophis sample (WII-ADR3491) we amplified and sequenced fragments of 16s gene, using the primers 16Sar (5’-CGCCTGTTTATCAAAAACAT-3’) + 16Sbr (5’-CCGGTCTGAACTCAGATCACGT-3’) (Palumbi et al. 1991). Polymerase chain reaction (PCR) condition followed was initial denaturation at 95°C for five minutes, followed by 35 cycles of denaturation at 95°C for 35 sec, annealing at 55°C for 45 sec, and extension at 72°C for one minute. The final extension was at 72°C for 10 min.  Amplified PCR products were run on a 2% agarose gel and viewed under a UV transilluminator. The purified PCR product was sequenced directly in an Applied Biosystems Genetic Analyzer 3500 XL in both directions using BigDye v3.1 kit.

We manually checked bidirectional sequences using the CHROMAS v2.6.6 software (http://technelysium.com.au/wp/chromas/) and aligned using ClustalW (Thompson et al. 199) with default prior settings implemented in MEGA v7.1 (Kumar et al. 2016). For the protein-coding gene (cyt b), we checked for unexpected stop codons by translating the sequence to amino acids in MEGA v7.1 (Kumar et al. 2016). We used Naja atra and Protobothrops mucrosquamatus as outgroup for phylogenetic analyses of Bungarus and Ovophis, respectively. The newly generated sequences were aligned with the sequences downloaded from GenBank (Benson et al. 2009) (Appendix 1).

We performed maximum likelihood (ML) analyses using IQ-TREE (Nguyen et al. 2015), implemented in the web server version (http://iqtree.cibiv.univie.ac.at) (Trifinopoulos et al. 2016). For cyt b, the dataset was partitioned by codon positions using Modelfinder (Kalyaanamoorthy et al. 2017), to find the best-fit partitions and models of sequence evolution for each partition (Appendix 2). Support for internal branches was quantified using the bootstrap approximation (UFB 1000 pseudoreplicates) (Minh et al. 2013). Uncorrected P-distance was calculated in MEGA v7.1 (Kumar et al. 2016) with pairwise deletions of missing data and gaps.

 

Morphological character scoring and identification

Total length, snout-vent length, and tail length were measured with a thread and a metre tape in millimetre (mm). Morphometric measurements were taken with the help of a digital calliper (Mitutoyo TM) to the nearest 0.1 mm. Length and width of head scales were measured at the longest and the widest points of the respective scales. Morphological comparisons follow Das et al. (2021). All measurements were taken on the specimen’s right side and scales were counted on both sides for bilateral scales. Morphometric and meristic characters were examined with the help of the Olympus SZX10 microscope. Dorsal scale rows (DSR) were counted at one head length behind the head, at midbody, and at one head length anterior to the anal scute. Ventral scales were counted following Dowling (1951). Bilateral scale counts separated by a comma are given in left, and right order. Abbreviations used are WII-ADR: Wildlife Institute of India Abhijit Das Reptile collection.

 

 

RESULTS

 

Molecular phylogeny

Based on mitochondrial gene cyt b, the specimen of Bungarus from Northeastern India clustered with the type sequences of B. suzhenae with strong bootstrap value (100) (Figure 2). Uncorrected P-distance between the newly collected Indian samples of Bungarus and the type series of B. suzhenae from the type locality varied 0.015–0.019 (Table 1). The inferred ML tree based on cyt–b showed B. suzhenae as a sister lineage to the cluster containing B. caeruleus, B. ceylonicus, B. lividus, and B. sindarus with low bootstrap value (48) (Figure 2).

Based on 16s gene, the Ovophis sample from Arunachal Pradesh nested with the Ovophis zayuensis sequence from Xizang province, China (with strong bootstrap value, (97) (Figure 3) along with two other samples from Yunnan province, China and Chin state, Myanmar. These samples altogether form a sister relationship with the samples of O. zayuensis from Tongmai and Xizang provinces, China with low node support (Figure 3). Uncorrected P-distance between our newly collected samples of Ovophis and samples of O. zayuensis from China and Myanmar was minimal (Table 2).

 

Morphology

We confirmed the new krait specimen as Bungarus suzhenae based on the following set of diagnostic morphological characteristics (Chen et al. 2021): enlarged hexagonal vertebral scale rows present, dorsal scales in 15:15:15 rows, anterior chin shields are slightly larger than the posterior chin shields, dorsal body black with 34 white bands, ventral scales connected with the black bands of the dorsal body by small dark patches, the ventral surface uniform white, underside of tail white with tiny brown dots in the middle.

The two viper specimens we collected were confirmed as Ovophis zayuensis based on the following set of diagnostic morphological characteristics (Che et al. 2020): the third supralabial larger is than the fourth, the second supralabial is fused with the loreal, internasal scales are separated by two scales; the ventral scale count is 169–172, dorsal scale count is 23 or 25:23:19 rows, the subcaudal count of 34–48, mostly single and some are paired, anal scale is single.

 

Description of Bungarus suzhenae (WII-ADR1241) (Image 1,2)

An adult male specimen. Body nearly triangular in cross-section, broader at midbody, more tapering posteriorly. Head slightly distinct from the neck and longer than broad (head width/head length = 0.5); head dorsally depressed at the parietal region, frontal area flat, gradually sloping towards snout from prefrontals; loreal region concave; supralabial sloping towards outer margin; snout rounded in dorsal and lateral view; eyes with rounded pupil; external nares oval, smaller than eye diameter; nasal large and divided; prenasal touches first supralabial, rostral and internasal; postnasal touches first and second supralabial, preocular, internasal and prefrontal; postnasal-preocular suture short and straight; pre-ocular one, hexagonal on right side and pentagonal left side, bordered by second and third supralabials, postnasal, prefrontal and supraocular; internasals two, wider than long, in contact with rostral, nasals and pre-frontals; prefrontals large, slightly wider than long; internasal suture shorter than prefrontal suture length and not aligned with latter; frontal shield-shaped, pointing posteriorly, 1.3 times longer than wide, bordered by prefrontals, supraoculars and parietals; supraoculars small, 1.5 times longer than wide, in contact with preoculars, upper postoculars, prefrontals, frontal and parietals; parietals large, longer than broad, bordered by frontal, supraoculars, upper postoculars, anterior temporal and upper posterior temporal on each side, and three smalls nuchal scales on pos-terior margins, posterolateral margins of parietals bordered by one enlarged elongate scales that anteriorly contact upper posterior temporals on each side; postoculars two on each side; temporals 1+2; supralabial seven on both side, third and fourth touches eye, lower postocular touches by fourth and fifth supralabials on each side; anterior temporal bordered by two postoculars, fifth and sixth supralabial, parietal, and two posterior temporals; lower posterior temporal in contact with sixth and seventh supralabial; rostral wider than long and triangular; mental smaller than rostral, bordered by first pair of infralabials; infralabial seven on both sides, fourth largest; first pair of chin shields slightly larger than the second pair chin shields; first chin shield is in contact with four infralabials on the right side and three infralabials on the left side; second chin shields contact with fourth infralabial on both sides, posteriorly bordered by three scales on the right side and fourth scales on the left side. Dorsal scales on body and tail smooth, no keel and apical pit; vertebral scale row enlarged and hexagonal; dorsal scale rows 15:15:15; ventrals 219; anal plate single; subcaudals 57, undivided; terminal scute conical; snout-vent length 664 mm and tail length 109 mm. Details of the morphometric and meristic characters of the specimen (WII-ADR1241) are given in (Table 3).

The hemipenes of the newly reported specimen of B. suzhenae from India agree with the description provided by Chen et al. (2021). Hemipenis can be divided into three distal calyculate region, middle spinose region, and basal region. The top of the hemipenis is slightly bilobed. The calyculate region is covered with well-developed small calyces. The spinose zone is covered with fang-shaped large keratinized spines and the basal region is covered with tiny spines; the basal region is nearly smooth towards the proximal end.

Dorsally head, body, and tail are uniformly dark brown. The rostral upper half is dark brown, the lower half (below the level of the lower edge of external nares) white; the nasals are also white below the level of the lower edge of external nares; supralabials are partly dark brown on the upper part and lower part (more than half of the scales) white; 34 narrow white cross bands on the dorsal body; bands widening on the flank before meeting the ventrals; a dark spot present at the junctions where the white bands meet the ventrals; white bands 5–7, 13, 15, 18–26, 28, and 31–33 are broad at mid dorsum with a central dark spot forming a white semicircle; white band 27 is incomplete; black bands wide and not intruding to venter; 13 white cross bands on the tail; ventrally white including infralabials, mental, chin shields, gular scales, ventrals, and subcaudals; the edge of the ventrals with slightly dark brown patch; tail ventrally white with tiny brown dots in the middle. The specimen is dorsally dark brown with white transverse bands on the body and tail. Interstitial skin white. Ventrally white, subcaudals with tiny brown dots in the middle (Image 2). Difference between the newly reported specimen and original description of B. suzhenae: Notable difference between the type specimen (based on the original description) and our newly reported specimen (WII-ADR1241) is a lower number of ventrals in the new specimen (219 vs. 220–229), the higher number of subcaudals in the new specimen (57 vs. 51–54).

This newly reported specimen was encountered along JessamiMeluri road (25.63930N, 94.57910E, elevation 599 m), at the Nagaland-Manipur border ca. 6 km from Meluri, Nagaland towards Jessami, Manipur. This individual was found while it was crossing the road at around 2100 h on 20 October 2012. The Tiji River flows ca. 20 m away from the collection locality. A roadkill Bungarus niger was also recorded 100 m away from the location of B. suzhenae indicating their sympatric occurrence. The surrounding landscape was mostly with secondary vegetation with extensive jhum cultivation areas. While photographing, the snake showed head-hiding behaviour and bit the snake bag in defence.

 

Description of Ovophis zayuensis (WII-ADR3316 and WII-ADR3491) (Image 3,4) 

Both the specimens are adult females. Specimens are in good condition, latero-ventral incision between ventral scale 99–102 in WII-ADR3316 and between ventral scale 66–70 in WII-ADR3491. Head triangular, body stout and the dorsal scales are strongly keeled, ventrally plain pale yellow or orangish in color with no patterns. There are faint black blotches on the anterior dorsal region of the body and the blotches are dark on the posterior dorsal region. After preservation, the specimen’s colour changed into smokey grey with black blotches on the dorsal side. Dorsally head is completely covered with small scales. The head length ranges 34.2–37.9 mm, the head width is 23.1–25 mm, the eye diameter is 2.5–2.6 mm, the eye-to-nostril distance is 5.5–7.3 mm, and the snout length is 10.1–11.3 mm. No supralabials touching the eye, one preocular and three postocular present. Nasal single and undivided in both specimens. There is no distinguishable temporal, parietal, and frontal scales present. The second supralabial is fused with the loreal pit.  The total length of the specimens is 826–900 mm, SVL 699–746 mm, tail length 127–154 mm, ventral scales 166–167 mm, anal scale single and undivided, subcaudals 46–49 with 5–6 paired. WII-ADR3491 has nine supralabials on the left, third is the largest measuring  4.3 mm; eight supralabials on the right, third is the largest measuring 3.6 mm. Infralabials 10 on the left, the fourth is the largest measuring  3.2 mm; 11 infralabials on the right and the sixth is the largest measuring 2.8 mm, dorsal scale rows 25:23:19. WII-ADR3316 has nine supralabials on both sides, third is the largest measuring 3.7 mm. Infralabials nine on both sides, on the right sixth is the largest measuring 3.5 mm, and on the left, fifth is the largest measuring 3.3 mm, dorsal scales rows 23:23:19.

The individual WII-ADR3491 was recorded at Dri River bank (28.80560N, 95.93210E, elevation 1,288 m), near Etabe village (~2.5 km north-east from Anini), Dibang Valley district, Arunachal Pradesh. This individual was found while it was moving on the ground along the edge of a first-order stream on 16 August 2022 at 2000 h. The stream edge was covered with small to moderate-sized boulders covered with moss and leaf litter, fern, and other vegetation. The recorded locality was ~100 m from the Dri river near Etabe village. Along the stream, we observed a few anuran species such as Xenophrys sp. and Amolops beibengensis on the vegetation and also an unidentified rodent species. The second individual WII-ADR3316 was collected from the newly constructed Chigu Pani road, Anini, Dibang Valley (29.01820N, 95.97550E, elevation 1,800 m). The habitat in this region was disturbed due to the recent construction of roads, small streams were blocked by retaining walls. It was found perched in a bush 50 cm above the ground around 1900 h between the walls, blocking the stream. There was a mild yet constant drizzle throughout the night. Many Amolops froglets were found near the blocked stream and on the road. This individual was a bit aggressive while trying to bag, it puffed up, flattened its body, and struck the snake hook. The next day around 2000 h on the same road, close to a small stream two other individuals of O. zayuensis male and female were spotted crossing the road from both sides. The blocked streams were seen to be harbouring tadpoles of multiple species of frogs including Amolops sp. and Megophrys sp. These two snakes were removed from the roadside and released by the stream.

 

 

DISCUSSION

 

There are over 400 known species of snakes in India (Uetz et al. 2024), where 1.2 million snakebite deaths were recorded during the years 2000–2019 (Suraweera et al. 2020). The Mountain Pit Viper Ovophis monticola Günther, 1864 has recently shown to cause medically significant envenoming (Ralph et al. 2023) while the Bungarus species have always been among the big four snakes of medical importance from India (Simpson & Norris 2007; Suraweera et al. 2020; Sunagar et al. 2021). Bungarus suzhenae and Ovophis zayuensis are two venomous snakes being reported for the first time from India, from the border of Nagaland-Manipur and Arunachal Pradesh, respectively.

Among the 18 known species of Bungarus, the maximum diversity is congregated in southeastern Asia. Until now eight species of Bungarus have been reported from India and are known for frequently causing fatal envenoming in the region (Suraweera et al. 2020; Sunagar et al. 2021). Despite several fatalities by other Bungarus species, the commercial antivenom is manufactured only against the most widespread congener B. caeruleus (Sunagar et al. 2021). This new finding reported here also indicates the lack of systematic studies on this group of snakes especially in northeastern India. Identification of Bungarus species is still a challenge which is evident by the recent discovery of B. suzhenae from China and Myanmar. Bungarus species with white cross bands—B. andamanensis Biswas & Sanyal, 1978; B. bungaroides Cantor, 1839; B. caeruleus Scheider, 1801; B. multicinctus Blyth, 1861; B. sindanus Boulenger, 1897; B. slowinskii Kuch, Kizirian, Nguyen, Lawson, Donnelly & Mebs, 2005; B. suzhenae Chen, Shi, Vogel, Ding & Shi, 2021; B. walli Wall, 1907; B. canidus Linnaeus, 1758; B. magnimaculatus Wall & Evans, 1901; and B. persicus Abtin, Nilson, Mobaraki, Hosseni & Dehgannejhad, 2014—are most complex and difficult to identify only based on external appearance (Leviton et al. 2003; Chen et al. 2021).

The genus Ovophis has eight known species globally (Malhotra et al. 2011), and now two species from India, namely: O. monticola and O. zayuensis. Though not commonly found in conflict with humans some species of Ovophis are capable of delivering a severely painful or life-threatening bite (Ralph et al. 2023). The venom composition of these less medically significant snakes and their clinical effects on humans still need to be studied (Tan et al. 2021). The current distribution range of O. zayuensis extends into the upper region of Dibang Valley, Arunachal Pradesh, Northeast India. However, it is important to carefully look into the distribution of O. zayuensis from the rest of Arunachal Pradesh. Being in the transition zone, the geographical range of these and other such species is likely to overlap in northeastern India. Therefore, proper identification of such venomous species and understanding of geographic distribution is crucial for general awareness, venom research as well as development of lifesaving antivenin.

Northeastern India is known to have Tibeto-Yunanese, Indo-Malayan, and its own unique faunal elements (Das 1996; Giri et al. 2019; Lalronunga et al. 2020; Vogel et al. 2020; Deepak et al. 2021; Das et al. 2021). The new record of B. suzhenae from India indicates the possibility of the occurrence of other Bungarus species reported from Myanmar such as B. magnimaculatus, B. wanghaotingi, and B. flaviceps. Therefore, a systematic study is necessary across northeastern India to document the diversity of medically important snakes. With the present report of B. suzhenae, the number of Bungarus species in India increases to nine, and the distribution range of B. suzhenae is extended by ca. 321 km north-west from the nearest known locality, Yingjiang County, Yunnan Province, China. The geographical area of occurrence of B. suzhenae is currently ca. 38,000 km2 within China, Myanmar, and northeastern India. In earlier documentation, other Bungarus species such as B. bungaroides, B. fasciatus, and B. niger were reported from Nagaland (Ao et al. 2004; Dasgupta & Raha 2006; Das 2018). Singh (1995) and Sinate et al. 2021 also reported B. fasciatus and B. niger from Manipur respectively. Since the present specimen of B. suzhenae was encountered at the Nagaland-Manipur border, there is a possibility of the occurrence of the species in other parts of Manipur. In a herpetofaunal biodiversity report of Nagaland, Grewal et al. (2011) provided a photograph of a road-killed unknown Bungarus sp. from Pungro-Shatuza road (nearest distance ca. 13 km from the present locality) which is similar to B. suzhenae (based on the lower number (<40) of unpaired white cross bands on body). Therefore, the distribution range of this medically important snake is likely to be wider than hitherto known in the region. Despite several reports of fatal snake bites by other species, commercial Indian antivenoms are only manufactured against Daboia russelii, Naja naja, Echis carinatus, and Bungarus caeruleus. Hence, it is important to recognise other medically important Indian snake species as it also has great significance in the field of snake antivenom design and manufacture (Simpson & Norris 2007).

 

Table 1. Uncorrected P-distances (%) of Bungarus species based on the cyt b sequences.

 

1

2

3

4

5

6

1. Bungarus suzhenae (India)

 

 

 

 

 

 

2. B. bungaroides (AY973270)

0.188

 

 

 

 

 

3. B. caeruleus (AJ749305)

0.135

0.189

 

 

 

 

4. B. candidus (AJ749329, AJ749339, AJ749341, MN165133)

0.098–0.101

0.167–0.17

0.139–0.142

0.003–0.008

 

 

5. B. ceylonicus (KC347457)

0.144

0.201

0.101

0.151

 

 

6. B. fasciatus (AJ749350, MW596457)

0.147–0.154

0.171–0.183

0.152–0.154

0.152–0.154

0.174–0.176

0.041

7. B. lividus (MW596472)

0.137

0.201

0.114

0.144

0.111

0.169

8. B. multicinctus (AJ749344, MN165136, MN165137, MN165139, MN165138, MN165135)

0.086–0.090

0.169–0.171

0.143–0.145

0.026–0.031

0.147–0.148

0.153–0.154

9. B. niger (AJ749304, MW596473)

0.091–0.095

0.177–0.181

0.135–0.14

0.101–0.106

0.144–0.146

0.152

10. B. sindanus (AJ749346)

0.145

0.186

0.129

0.136

0.134

0.166

11. B. slowinskii (AJ749306)

0.176

0.087

0.190

0.179

0.199

0.174

12. B. suzhenae (AJ749435, MN165142, MN165140, MN165141, MN165143)

0.015–0.019

0.185–0.187

0.133–0.136

0.096–0.1

0.138–0.142

0.151–0.156

13. B. wanghaotingi (MN165144, KY952766, MN165146, MN165145, MN165132, AJ749336, AJ749308, MN165134, AJ749309, MN165131, AJ749331, AJ749337)

0.092–0.1

0.163–0.177

0.136–0.144

0.018–0.024

0.142–0.151

0.148–0.161

Table 1. continue…

7

8

9

10

11

12

0.136–0.139

0–0.006

 

 

 

 

0.14–0.143

0.102–0.104

 

 

 

 

0.142

0.130

0.129

 

 

 

0.197

0.170

0.171

0.183

 

 

0.135–0.140

0.087–0.091

0.091–0.095

0.141–0.144

0.171–0.176

0.002–0.007

0.134–0.139

0.026–0.03

0.096–0.102

0.126–0.137

0.164–0.175

0.087–0.097

 

Table 2. Uncorrected P-distances (%) of Ovophis species based on the 16s gene sequences.

 

1

2

3

4

5

6

1. Ovophis zayuensis (India)

 

 

 

 

 

 

2. O. zayuensis (HQ325109, HQ325111, HQ325118, HQ325089, MK193194, MK193195)

0.004–0.019

0.0–0.013

 

 

 

 

3. O. monticola (HQ326117, HQ325121, HQ325078, MG995792)

0.05–0.062

0.039–0.057

0.004–0.017

 

 

 

4. O. makazayazaya (HQ325107)

0.056

0.039–0.055

0.05–0.054

 

 

 

5. O. tonkinensis (HQ325096, HQ325070)

0.031–0.032

0.026–0.032

0.049–0.056

0.03–0.034

0.004

 

6. O. okinavensis  (AB175670)

0.056

0.035–0.046

0.057–0.059

0.054

0.052–0.053

 

7. O. convictus (HQ325082, HQ325083)

0.046–0.052

0.032–0.05

0.041–0.059

0.05–0.052

0.042–0.043

0.05–0.056

 

 

Table 3. Morphometric and meristic data of newly collected specimens of Bungarus suzhenae. All the measurements are given in millimetre. Bilateral characters are given in left, and right order separated by comma. “N” denotes sample size, “-“ indicates data not provided.

Voucher ID

WIIADR1241

Chen et al. (2021)

Sex

male

male (N = 3)

female

Snout- vent length (SVL)

664

620–1140

1310

Tail length (TL)

109

109–180

-

Head length (HL)

21.6

21–39

30.2

Head width (HW)

10.7

12.3–15.5

19.4

Head height (HH)

6.5

8.7–12.8

14.2

Eye diameter

2.3

9.3–10.5

14.6

Ventrals

219

220–229

222

Subcaudals

57

51–54

11+

Anal plate

single

single

single

Dorsal scale row

15:15:15

15:15:15

15:15:15

Supralabial

7,7

7,7

7,7

Infralabial

7,7

7,7

7,7

Preocular

1,1

1,1

1,1

Postocular

2,2

1–2

2

Temporal (anterior + posterior)

1+2

1+2

1+2

Nasal

divided

divided

divided

No. of white bands (body + tail)

34+13

26–38+9–12

34+3

 

 

Table 4. Morphometric and meristic data of newly collected specimens of Ovophis zayuensis. All the measurements are given in millimetre.

Voucher ID

WII-ADR3491

WII-ADR3316

Sex

female

female

Snout- vent length (SVL)

746

699

Tail length (TL)

154

127

Head length (HL)

37.9

34.2

Head width (HW)

25

23.1

Head height (HH)

14.3

14.2

Eye diameter

2.6

2.5

Ventrals

167

166

Subcaudals

49

46 (missing tip)

Anal plate

single

single

Dorsal scale row

25:23:19

23:23:19

Supralabial

9,8

9,9

Infralabial

10,11

9,9

Preocular

1,1

1,1

Postocular

3,3

3,3

Nasal

undivided

undivided

 

 

Identification key to the Bungarus species of northeastern India

Crossbands on body and tail absent ......................................................................................... 1

1A. Mid-dorsal scale distinctly enlarged .................................................................................... Bungarus niger

1B. Mid-dorsal scale not enlarged .............................................................................................. Bungarus lividus

Crossbands present ..................................................................................................................... 2

2A. Crossbands yellow and black ................................................................................................. Bungarus fasciatus

2B. Crossbands white .................................................................................................................... 3

3A. White crossbands not paired, 26–38 in number ................................................................ Bungarus suzhenae

3B. White crossbands not paired, 46–60 in number ................................................................. Bungarus bungaroides

 

Identification key to the Ovophis species from India

Ventral scales

A. 135–154 .................................................................................................................................... O. monticola

B. 158–176  .................................................................................................................................. O. zayuensis

 

 

For figures & images - - click here for full PDF

 

 

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Appendix 1. Details of the GenBank sequences used in this study. The sequence of Bungarus suzhenae and Ovophis zayuensis generated in this study is given in bold.

Taxa

Voucher numbers

 Locality

AccessionNo. (cytb)

Reference

B. bungaroides

KIZ 98R0186

Medog, Medog County, Tibet, China

AY973270

Kuch et al. 2005

B. caeruleus

UK H7

Pakistan

AJ749305

Kuch 2007

B. candidus

UK B39

Vicinity of Jambah, Losarang District, Indramayu Regency, West Java, Indonesia

AJ749329

Kuch 2007

B. candidus

UK BcBa

Bali Island, Indonesia

AJ749339

Kuch 2007

B. candidus

UK B21

Vicinity of Jambah, Losarang District, Indramayu Regency, West Java, Indonesia

AJ749341

Kuch 2007

B. candidus

SYNU R180411

Endau Rompin National Park, Taman Negara, Johor, Malaysia

MN165133

Chen et al. 2021

B. ceylonicus

RS-135

Sri Lanka

KC347457

Pyron et al. 2013

B. fasciatus

UK B24

Vicinity of Jolo, Central Java, Indonesia

AJ749350

Kuch 2007

B. fasciatus

MZMU978

Mizoram, India

MW596475

Biakzuala et al. 2021

B. lividus

YSR187

Baridua, Meghalaya, India

MW596472

Biakzuala et al. 2021

B. multicinctus

CIB104228

Nanning Zoo, Guangxi, China (locality uncertain)

MN165137

Chen et al. 2021

B. multicinctus

UK Bm1

Zhejiang, China

AJ749344

Kuch 2007

B. multicinctus

CIB DL18090209

Wuyi Moutain, Fujian, China

MN165138

Chen et al. 2021

B. multicinctus

CIB DL18090210

Wuyi Moutain, Fujian, China

MN165139

Chen et al. 2021

B. multicinctus

CIB93923

Guangxi, China

MN165136

Chen et al. 2021

B. multicinctus

SYNU R180305

Haikou, Hainan, China

MN165135

Chen et al. 2021

B. niger

Bnig

Nepal

AJ749304

Kuch 2007

B. niger

MZMU975

Mizoram, India

MW596473

Biakzuala et al. 2021

B. sindanus

Bsin1

Pakistan

AJ749346

Kuch 2007

B. slowinskii

IEBR 1172

Na Hau Commune, Van Yen District, Yen Bai,Vietnam

AJ749306

Kuch 2007

B. suzhenae

CAS 221526

Naung Mon Township, Rabaw Kachin State, Burma (Myanmar)

AJ749345

Kuch 2007

B. suzhenae

CIB116088

Yingjiang, Yunnan, China

MN165140

Chen et al. 2021

B. suzhenae

CIB116089

Yingjiang, Yunnan, China

MN165141

Chen et al. 2021

B. suzhenae

CIB116090

Yingjiang, Yunnan, China

MN165142

Chen et al. 2021

B. suzhenae

CIB116091

Yingjiang, Yunnan, China

MN165143

Chen et al. 2021

B. suzhenae

WIIADR1241

Nagaland-Manipur border, India

PP808595

This study

B. wanghaotingi

ROM 35250

Quang Thanh, Cao Bang, Vietnam

AJ749308

Kuch 2007

B. wanghaotingi

ROM 35256

Chi Linh, Hia Duong, Vietnam

AJ749309

Kuch 2007

B. wanghaotingi

UK BT6

Nakhon Si Thammarat, Thailand

AJ749331

Kuch 2007

B. wanghaotingi

UK BT8

Nakhon Si Thammarat, Thailand

AJ749336

Kuch 2007

B. wanghaotingi

FMNH 255259

Hin Nam No National Biodiversity Conservation Area, Boualapha District, Khammouan, Laos

AJ749337

Kuch 2007

B. wanghaotingi

FMNH 255260

Nam Et Phou Louey Mountain, Phou Louey National Biodiversity, Conservation Area, Vieng Tong, Huaphahn, Laos

AJ749338

Kuch 2007

B. wanghaotingi

CIB FCDZ20170806

Dongzhong, Fangchenggang, Guangxi, China

MN165131

Chen et al. 2021

B. wanghaotingi

CIB104227

Beiliu, Guangxi, China

MN165134

Chen et al. 2021

B. wanghaotingi

SYNU R170408

Bang Lang National Park, Yala, Thailand

MN165132

Chen et al. 2021

B. wanghaotingi

CIB ML20170801

Menglun, Yunnan, China

MN165145

Chen et al. 2021

B. wanghaotingi

CIB MLMY20170801

Mengla, Yunnan, China

MN165144

Chen et al. 2021

B. wanghaotingi

JK20181101

Jinghong, Yunnan, China

MN165146

Chen et al. 2021

B. wanghaotingi

GP 3375

Xishuangbanna, Yunnan Province, China

KY952766

Xie et al. 2018

Naja atra

-

China

DQ343648

Yan et al. 2008

O. zayuensis

WII-ADR3491

Dibang Valley district, Arunachal Pradesh, India

PP794643

This study

O. zayuensis

 

GP90

Xizang Autonomous Region, China

HQ325109

Malhotra et al. 2011

O. zayuensis

 

GP92

Yunnan Province, China

HQ325111

Malhotra et al. 2011

O. zayuensis

CAS 233203

Chin State, Myanmar

HQ325118

Malhotra et al. 2011

O. zayuensis

CHS099

Tibet, China

MK193914

Li, J. N et al. 2020

O. zayuensis

CHS101

Tibet, China

MK193915

Li, J. N et al. 2020

O. zayuensis

GP89

Xizang Autonomous Region, China

HQ325089

Malhotra et al. 2011

O. tonkinensis

MVZ 226627

Vinh Phu Province, N Viet Nam

HQ325096

Malhotra et al. 2011

O. tonkinensis

B581

China

HQ325070

Malhotra et al. 2011

O.monticola makazayazaya

GP24

Yunnan Province, China

HQ325107

 

Malhotra et al. 2011

O. monticola

CAS 224424

Kachin State, Myanmar

HQ325117

Malhotra et al. 2011

O. monticola

V16

North East India

MG995792

 

O. monticola

 

CAS 234866

Chin State, Myanmar

                                                      

HQ325121

Malhotra et al. 2011

O. monticola

 

ZMB 70216

Gandaki Zone, W Nepal

HQ325078

Malhotra et al. 2011

O. okinavensis

 

-

-                                                           

AB175670

Dong & Kumazawa 2005

 

O. convictus

B629

Pulau Langkawi, West Malaysia

HQ325083

Malhotra et al. 2011

O. convictus

B628

Pulau Langkawi, West Malaysia

HQ325082

Malhotra et al. 2011

Protobothrops mucrosquamatus

B106

Vin Phuc Province, N Viet Nam

AY294271

Herrmann et al. 2004