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Bungarus multicinctus (B. multicinctus) is one of the top ten venomous snakes in China, ranking first in lethality at 26.9-33.3%. However, to our knowledge, no forensic autopsy-related cases of death from B. multicinctus bite poisoning have been reported. There are surprisingly few reported cases of death from poisoning by other species of neurotoxic snakes. Neurotoxic snake venom is often highly toxic, and death can quickly occur when bitten in the wild if victims are not taken to a doctor in time. We presented a case of an adult female in Fujian Province of China who was bitten by a poisonous snake while digging for bamboo shoots in the mountains and died from the bite of B. multicinctus confirmed by enzyme-linked immunosorbent assays (ELISA) results. The autopsy's results, histopathological findings, and ELISA results reported here can be helpful for future forensic practice in B. multicinctus venom poisoning; we also briefly review the pathological changes of neurotoxin poisoning, which may be useful in other types of neurotoxin snake venom poisoning.

Bungarus candidus (Linnaeus, 1758) — Erroneous. Malayan Krait Singapore records. Bungarus candidus —Sharma, 1973: 235.—Chou, 1986: 8, 13.—F.L.K. Lim & Lee, 1989: 116.— Gopalakrishnakone, 1990: 3.—K.K.P. Lim & L.M. Chou, 1990: 54.—F.L.K. Lim, 1991: 74.— K.K.P. Lim & F.L.K. Lim, 1992: 87, 148.—L.M. Chou et al., 1994: 105.— David & Vogel, 1996: 143.— Manthey & Grossmann, 1997: 416.—Chan-ard et al., 1999: 37.— Iskandar & Colijn, 2001: 119.—K.P. Lim & F.L.K. Lim, 2002: 148.— de Lang & Vogel, 2005: 259.—N. Baker & K.P. Lim, 2008: 169.— Das, 2010: 313.— Chanhome et al., 2011: 314.—P.K.L. Ng et al., 2011: 302.—N. Baker & K.P. Lim, 2012: 169.— Das, 2012a: 88.— Mohammadi et al., 2014: 21.—Wallach et al., 2014: 128.—Chan-ard et al., 2015: 250.— de Lang, 2017: 238.— Cox et al., 2018: 50.— Das, 2018: 105.—Y. Xie et al., 2018: 109.— Grahadi et al., 2022: 1047, 1049, 1052. “Malayan krait”—Mong & H.H. Tan, 2016: 266. Remarks. All of the publications listed in the chresynonymy report B. candidus as occurring in Singapore but none list any material or specific source for the record. We have also not been able to find any earlier references to B. candidus from Singapore and no specimens exist in museum collections. We suspect inclusion of B. candidus into Singapore’s herpetofauna checklist may stem from confusion with B. fasciatus. A piece of shed skin found at Pulau Tekong in 2002 is said to be of B. candidus, but was not confirmed (K.K.P. Lim unpub. data). Bungarus candidus is distributed from Thailand, Cambodia, and Vietnam south to Peninsular Malaysia, and Bawean, Bali, Java, Karimunjawa, and Sumatra in Indonesia (Wallach et al. 2014). LKCNHM & NHMUK Museum specimens. Pulau Tekong: ZRC.2.5485 (02-Jul-2002). Additional Singapore museum specimens. No specimens., Published as part of Figueroa, Alex, Low, Martyn E. Y. & Lim, Kelvin K. P., 2023, Singapore's herpetofauna: updated and annotated checklist, history, conservation, and distribution, pp. 1-378 in Zootaxa 5287 (1) on page 273, DOI: 10.11646/zootaxa.5287.1.1, http://zenodo.org/record/7960319, {"references":["Gopalakrishnakone, P. (1990) A Colour Guide to Dangerous Animals. Singapore University Press, Singapore, 156 pp.","Lim, K. K. P. & Lim, F. L. K. (1992) A Guide to the Amphibians and Reptiles of Singapore. Singapore Science Centre, Singapore, 160 pp.","Chou L. M., Ng, P. K. L. & Lim, K. K. P. (1994) Animalia. In: Wee, Y. C. & Ng, P. K. L. (Eds.), A First Look at Biodiversity in Singapore. National Council on the Environment, Singapore, pp. 70 - 106.","David, P. & Vogel, G. (1996) The Snakes of Sumatra: An Annotated Checklist and Key with Natural History Notes. Edition Chimaira, Frankfurt-am-Main, 260 pp.","Manthey, U. & Grossmann, W. (1997) Amphibien und Reptilian Sudostasien. Natur und Tier, M ¸ nster, 512 pp.","Iskandar, D. T. & Colijn, E. (2001) A Checklist of Southeast Asian and New Guinean Reptiles. Part I. Serpentes. Biodiversity Conservation Project (Indonesian Institute of Sciences - Japan International Cooperation Agency - The Ministry of Forestry). The Gibbon Foundation and Institute of Technology, Bandung, 195 pp.","Lim, K. P. & Lim, F. L. K. (2002) A Guide to the Amphibians and Reptiles of Singapore. Revised Edition. Singapore Science Centre, Singapore, 160 pp.","de Lang, R. & Vogel, G. (2005) The Snakes of Sulawesi: A Field Guide to the Land Snakes of Sulawesi with Identification Keys. Edition Chimaira, Frankfurt am Main, 312 pp.","Das, I. (2010) A Field Guide to the Reptiles of Thailand and South-East Asia. New Holland, London, 376 pp.","Chanhome, L., Cox, M. J., Vasaruchapong, T., Chaiyabutr, N. & Sitprija, V. (2011) Characterization of venomous snakes of Thailand. Asian Biomedicine, 5 (3), 311 - 328.","Das, I. (2012 a) A Naturalist's Guide to the Snakes of South-East Asia. John Beaufoy Publishing, Oxford, 160 pp.","Mohammadi, S., Kluever, B. M., Tamashiro, T., Amano, Y. & Hill III, J. G. (2014) Short note spatial and Thermal observations of a Malayan Krait (Bungarus candidus) From Thailand. Tropical Natural History, 14 (1), 21 - 26.","de Lang, R. (2017) The Snakes of Java, Bali and Surrounding Islands. Edition Chimaira, Frankfurt am Main, 435 pp.","Cox, M. J., Hoover, M. F., Chanhome, L., Thirakhupt, K. & Pongcharoen, C. (2018) A Field Guide to the Venomous Snakes of Mainland ASEAN Nations. Parbpim Limited Partnership, Bangkok, ix + 83 pp.","Das, I. (2018) A Naturalist's Guide to the Snakes of Southeast Asia. 2 nd Edition. John Beaufoy Publishing, Oxford, 176 pp.","Grahadi, R., Fatchiyah, F. & Kurniawan, N. (2022) Virtual prediction of potential immunogenic epitope of candoxin protein from Malayan krait (Bungarus candidus) venom. Journal of Pharmacy & Pharmacognosy Research, 10 (6), 1046 - 1057. https: // doi. org / 10.56499 / jppres 22.1469 _ 10.6.1046"]}

Bungarus flaviceps Reinhardt, 1843 — Erroneous. Red-headed Krait Singapore records. Bungarus flaviceps flaviceps — Chanhome et al., 2011: 315.— Cox et al., 2018: 52. Remarks. Chanhome et al. (2011) listed Singapore as part of the distribution for B. flaviceps, but we are unsure where they acquired the record from since there are no publications reporting it from Singapore. Cox et al. (2018) likely reference Chanhome et al. (2011). Bungarus flaviceps ranges from southern Myanmar east to southern Vietnam, and south to Peninsular Malaysia, Borneo, and Banka, Belitung, Java, Nias, and Sumatra in Indonesia (Wallach et al. 2014). LKCNHM & NHMUK Museum specimens. No specimens. Additional Singapore museum specimens. No specimens. Genus Hydrophis Latreille in Sonnini & Latreille, 1801 (11 species), Published as part of Figueroa, Alex, Low, Martyn E. Y. & Lim, Kelvin K. P., 2023, Singapore's herpetofauna: updated and annotated checklist, history, conservation, and distribution, pp. 1-378 in Zootaxa 5287 (1) on pages 273-274, DOI: 10.11646/zootaxa.5287.1.1, http://zenodo.org/record/7960319, {"references":["Chanhome, L., Cox, M. J., Vasaruchapong, T., Chaiyabutr, N. & Sitprija, V. (2011) Characterization of venomous snakes of Thailand. Asian Biomedicine, 5 (3), 311 - 328.","Cox, M. J., Hoover, M. F., Chanhome, L., Thirakhupt, K. & Pongcharoen, C. (2018) A Field Guide to the Venomous Snakes of Mainland ASEAN Nations. Parbpim Limited Partnership, Bangkok, ix + 83 pp."]}

Bungarus fasciatus (Schneider, 1801) — Native. Pseudoboa Fasciata Schneider, 1801: 283–284. Syntypes (2): ZMB 2771–72, by original designation. Type locality: “Indien” (= India); later restricted to “Mansoor Cottah, Bengal, India ” by Russell (1801: 53). Banded Krait (Figure 21F) Singapore records. Bungarus fasciatus —Peters, 1859: 271.— Dennys, 1880a: 3.— Blanford, 1881: 216.— Davison, 1892: 89, 92.— Flower, 1896: 893.— Flower, 1899: 689.—Ridley, 1899: 209.—Sworder, 1923: 71.— Sworder, 1924a: 19, 20 (Pulau Ayer Merbau).—Sworder, 1924b: 22 (Tanglin [= TF]).— Buddle, 1929: 21 (Sembawang Naval Base [= SML]).—Smith, 1930: 67 (“near Singapore town”).—D.S. Johnson, 1964: 27.—Sharma, 1973: 235.—F.L.K. Lim, 1984: 17.—F.L.K. Lim & M.T.-M. Lee, 1989: 115.— Gopalakrishnakone, 1990: 3.—K.K.P. Lim & L.M. Chou, 1990: 54.—F.L.K. Lim, 1991: 74.—D.S. Johnson, 1992: 38.—K.K.P. Lim & F.L.K. Lim, 1992: 86, 148.—L.M. Chou et al., 1994: 105.— David & Vogel, 1996: 144.— Manthey & Grossmann, 1997: 417.— Cox et al., 1998: 30.—Chan-ard et al., 1999: 37.—E.K. Chua, 2000: 118 (Pulau Ubin).— Iskandar & Colijn, 2001: 120.—K.P. Lim & F.L.K. Lim, 2002: 148.—N. Baker & K.P. Lim, 2008: 115, 161.—K.K.P. Lim et al., 2008: 166, 265 (Khatib Bongsu; Lim Chu Kang; Pulau Ubin; Pulau Tekong; Sungei Buloh [SBWR]).—P.K.L. Ng et al., 2008: 168.— Das, 2010: 313.— Chanhome et al., 2011: 315.— P.K.L. Ng et al., 2011: 302.—N. Baker & K.P. Lim, 2012: 115, 161.—de Chambrier et al., 2012: 45.— Das, 2012a: 89.—H.H. Tan, 2013: 2 (Pulau Ubin).—Wallach et al., 2014: 128.—Chan-ard et al., 2015: 250.—K.K.P. Lim et al., 2016: 184 (Pulau Tekong).— Ahsan & Rahman, 2017: 9907.— de Lang, 2017: 245.— Cox et al., 2018: 51.— Das, 2018: 105.—H.Y. Lim, 2019: 128–129 (Chek Jawa Wetlands [PU]).— Charlton, 2020: 224. Bungarus fasciatus fasciatus — de Haas, 1950: 597. “banded krait”—Mong & H.H. Tan, 2016: 266. Remarks. Throughout its range, B. fasciatus inhabits a diverse range of habitats including montane areas (Stuebing et al. 2014). However, in Singapore, B. fasciatus appears restricted mainly to coastal habitats (Baker & Lim 2012) with most observations coming from Pulau Ubin. Bungarus fasciatus was first reported from Singapore by Peters (1859) who listed a specimen at ZMB collected by Jagor. Subsequently, Blanford (1881) examined specimens at the Raffles Museum, and Davison (1892) noted an additional specimen that was mutilated and deposited at the Raffles Museum around that time. No other specimens were recorded until Sworder (1924a) observed one at Pulau Ayer Merbau, and mentioned Smith killed one near Tanglin in 1901 (Sworder 1924b). Shortly after, Buddle (1929) captured one at SNB and killed one near to town (Smith 1930). Bungarus fasciatus was then next reported 61 years later (Table 2) by Lim (1991) who related how a specimen he received was caught in a fishing net along a riverbank. The only other three published observations include one photographed in mangroves at PT in May 2006 (Lim et al. 2016), one crossing a road near mangroves at PU on 18 October 2012 (Tan 2013), and one seemingly foraging in mud lobster mounds on 4 August 2019 at PU (Lim 2019). Figure 21F illustrates one that was rescued from a contractor dormitory next to Ubin Fruit Orchard on PU on 12 January 2018 (I.S. Law pers. comm.). Lim et al. (2008) reported B. fasciatus from KBNP, LCK, PU, and SBWR, but did not provide any specific data, except for the LCK locality which was based on ZRC.2.4658 collected on 1 November 1999. Occurrence. Known mainly from coastal areas in Lim Chu Kang, PU, and PT. Uncommon. Singapore conservation status. Vulnerable. Conservation priority. Highest. IUCN conservation status. Least Concern [2013]. LKCNHM & NHMUK Museum specimens. Singapore (no locality): ZRC.2.6384– ZRC.2.6385 (no date), ZRC.2.6387 (1986); Lim Chu Kang Road [LKC]: ZRC.2.4658 (01-Nov-1999). Additional Singapore museum specimens. Singapore (no locality): NMW, ZMB; Jurong Road [= Tengah Forest]: BPBM. Singapore localities. Khatib Bongsu Nature Park—Lim Chu Kang—Pulau Ayer Merbau*—Pulau Tekong—Pulau Ubin—Sembcorp Marine Ltd.*—Sungei Buloh Wetland Reserve—Tyersall Forest*—Tengah Forest. Genus Calliophis Gray, 1835 (3 species) Calliophis Gray, 1835 [in 1832–1835]: pl. 86, fig. 1, caption (type species: Calliophis gracilis Gray, 1835 [in 1832–1835], by monotypy; gender masculine)., Published as part of Figueroa, Alex, Low, Martyn E. Y. & Lim, Kelvin K. P., 2023, Singapore's herpetofauna: updated and annotated checklist, history, conservation, and distribution, pp. 1-378 in Zootaxa 5287 (1) on pages 189-190, DOI: 10.11646/zootaxa.5287.1.1, http://zenodo.org/record/7960319, {"references":["Dennys, N. B. (1880 a) The snakes of Singapore I. Singapore Daily Times, 20 December 1880, 2 - 3.","Blanford, W. T. (1881) On a collection of reptiles and frogs chiefly from Singapore. Proceedings of the Zoological Society of London, 1881 (1), 215 - 226, pls. 20 - 21. https: // doi. org / 10.5962 / bhl. title. 101596","Davison, W. (1892) The snakes of Singapore. Singapore and Straits Directory, 1892, 87 - 93.","Flower, S. S. (1896) Notes on a collection of reptiles and batrachians made in the Malay Peninsula in 1895 - 96; with a list of the species recorded from that region. Proceedings of the Zoological Society of London, 1896 (4), 856 - 914, pls. 44 - 46.","Flower, S. S. (1899 b) Notes on a second collection of reptiles made in the Malay Peninsula and Siam, from November 1896 - September 1898, with a list of the species recorded from those countries. Proceedings of the Zoological Society of London, 1899 (4), 600 - 696, pls. 36 - 37.","Buddle, R. (1929) Snakes of Singapore Island. Kelly & Walsh, Limited, Singapore, 48 pp.","Johnson, D. S. (1964) An Introduction to the Natural History of Singapore. Rayirath (Raybooks) Publications, Singapore, x + 106 pp.","Lim, F. L. K. (1984) Reptile and amphibian encounters. In: Wee, Y. - C. (Ed.), Nature in an Urban Singapore. Proceedings of the Seminar on Nature in an Urban Singapore. Singapore Branch of the Malayan Nature Society, Singapore, pp. 17 - 19.","Gopalakrishnakone, P. (1990) A Colour Guide to Dangerous Animals. Singapore University Press, Singapore, 156 pp.","Johnson, D. S. (1992) Natural History of Singapore. Revised Edition. Tynron Press, Leicestershire, ix + 142 pp.","Lim, K. K. P. & Lim, F. L. K. (1992) A Guide to the Amphibians and Reptiles of Singapore. Singapore Science Centre, Singapore, 160 pp.","Chou L. M., Ng, P. K. L. & Lim, K. K. P. (1994) Animalia. In: Wee, Y. C. & Ng, P. K. L. (Eds.), A First Look at Biodiversity in Singapore. National Council on the Environment, Singapore, pp. 70 - 106.","David, P. & Vogel, G. (1996) The Snakes of Sumatra: An Annotated Checklist and Key with Natural History Notes. Edition Chimaira, Frankfurt-am-Main, 260 pp.","Manthey, U. & Grossmann, W. (1997) Amphibien und Reptilian Sudostasien. Natur und Tier, M ¸ nster, 512 pp.","Cox, M. J., van Dijk, P. P., Nabhitabhata, J. & Thirakhupt, K. (1998) A Photographic Guide toSnakes and other Reptiles of Peninsular Malaysia, Singapore and Thailand. New Holland Publishers, London, 144 pp.","Chua, E. K. (2000) Pulau Ubin. Ours to Treasure. Simply Green, Singapore, 140 pp.","Iskandar, D. T. & Colijn, E. (2001) A Checklist of Southeast Asian and New Guinean Reptiles. Part I. Serpentes. Biodiversity Conservation Project (Indonesian Institute of Sciences - Japan International Cooperation Agency - The Ministry of Forestry). The Gibbon Foundation and Institute of Technology, Bandung, 195 pp.","Lim, K. P. & Lim, F. L. K. (2002) A Guide to the Amphibians and Reptiles of Singapore. Revised Edition. Singapore Science Centre, Singapore, 160 pp.","Das, I. (2010) A Field Guide to the Reptiles of Thailand and South-East Asia. New Holland, London, 376 pp.","Chanhome, L., Cox, M. J., Vasaruchapong, T., Chaiyabutr, N. & Sitprija, V. (2011) Characterization of venomous snakes of Thailand. Asian Biomedicine, 5 (3), 311 - 328.","Chambrier, A. de, Binh, T. T. & Scholz, T. (2012) Ophiotaenia bungari n. sp. (Cestoda), a parasite of Bungarus fasciatus (Schneider) (Ophidia: Elapidae) from Vietnam, with comments on relative ovarian size as a new and potentially useful diagnostic character for proteocephalidean tapeworms. Systematic Parasitology, 81 (1), 39 - 50. https: // doi. org / 10.1007 / s 11230 - 011 - 9320 - 0","Das, I. (2012 a) A Naturalist's Guide to the Snakes of South-East Asia. John Beaufoy Publishing, Oxford, 160 pp.","Lim, K. K. P., Chua, M. A. H. & Lim, N. T. - L. (2016) Freshwater fishes, terrestrial herpetofauna and mammals of Pulau Tekong, Singapore. Nature in Singapore, 9, 165 - 198.","Ahsan, M. F. & Rahman, M. M. (2017) Status, distribution and threats of kraits (Squamata: Elapidae: Bungarus) in Bangladesh. Journal of Threatened Taxa, 9 (3), 9903 - 9910. https: // doi. org / 10.11609 / jott. 2929.9.3.9903 - 9910","de Lang, R. (2017) The Snakes of Java, Bali and Surrounding Islands. Edition Chimaira, Frankfurt am Main, 435 pp.","Cox, M. J., Hoover, M. F., Chanhome, L., Thirakhupt, K. & Pongcharoen, C. (2018) A Field Guide to the Venomous Snakes of Mainland ASEAN Nations. Parbpim Limited Partnership, Bangkok, ix + 83 pp.","Das, I. (2018) A Naturalist's Guide to the Snakes of Southeast Asia. 2 nd Edition. John Beaufoy Publishing, Oxford, 176 pp.","Charlton, T. (2020) A Guide to Snakes of Peninsular Malaysia and Singapore. Natural History Publications (Borneo) Sdn. Bhd., Kota Kinabalu, viii + 299 pp.","de Haas, C. P. J. (1950) Checklist of the snakes of the Indo-Australian Archipelago (Reptiles, Ophidia). Treubia, 20 (3), 511 - 625.","Baker, N. & Lim, K. P. (2012) Wild Animals of Singapore: A Photographic Guide to Mammals, Reptiles, Amphibians and Freshwater Fishes. Updated Edition. Draco Publishing and Distribution Pte Ltd and Nature Society, Singapore, 180 pp."]}

Inefficacy and associated risks of current antivenom has raised the need for alternative approaches of snakebite management. Aptamers are one such alternative which is being pursued for therapeutic interventions as well as for design of diagnostic kits due to its high specificity. Present study focussed on designing and validating nucleic acid aptamers against snake venom PLA2, a hydrolytic enzyme present in all venomous snakes. The aptamers were designed by adding nucleic acid chain on the surface of Daboxin P, a major PLA2 enzyme of Daboia russelii venom. Binding characteristics of the aptamers were confirmed by docking to Daboxin P as well as acidic and basic PLA2s from different snake species using in silico docking. The aptamers folded into different tertiary structures and bound to the active and Ca2+ binding site of PLA2 enzymes. Molecular dynamics simulation analysis of Daboxin P-aptamer complexes showed that the complexes were stable in an aqueous environment. The electrophoretic mobility shift assay further confirmed the binding of the synthetic aptamers to Daboxin P and other snake venom PLA2 enzymes. The aptamers inhibited the sPLA2 activity with an IC50 value ranging between 0.52 μM and 0.77 μM as well as the anticoagulant activity of Daboxin P. The aptamers could also inhibit the PLA2 activity of Echis carinatus crude venom and anti-coagulant activity of Bungarus caeruleus crude venom, members of big four snakes. However, the aptamers didn't inhibit fibrinogenolytic or proteolytic activity of big four venom as well as the coagulation and hemolytic activities. Thus, aptamers can be rationally designed to inhibit the biochemical and biological activities of snake venom proteins.

Bungarus multicinctus is one of the top ten venomous snakes in China, and its bite causes acute and severe diseases, but its pathophysiology remains poorly elucidated. Thus, an animal model of Bungarus multicinctus bite was established by intramuscular injection of 30μg/kg of Bungarus multicinctus venom, and then the serum metabolites were subsequently screened, identified and validated by ultra-performance liquid chromatography-quadrupole-time of flight-mass spectrometry (UPLC-Q-TOF-MS) methods to explore the potential biomakers and possible metabolic pathways. Untargeted metabolomics analysis showed that 36 and 38 endogenous metabolites levels changed in ESI+ and ESI-, respectively, KEGG pathway analysis showed that 5 metabolic pathways, including mineral absorption, central carbon metabolism in cancer, protein digestion and absorption, aminoacyl-tRNA biosynthesis and ABC transporters might be closely related to Bungarus multicinctus bite. Targeted metabolomics analysis showed that there were significant differences in serum D-proline, L-leucine and L-glutamine after Bungarus multicinctus bite (P < 0.05). In addition, receiver operating characteristic (ROC) analysis showed that the diagnostic efficiency of L-Glutamine was superior to other potential biomarkers and the AUC value was 0.944. Moreover, we found evidence for differences in the pathophysiology of glutamine between Bungarus multicinctus bite group and normal group, specifically with the content of glutamine synthetase (GS) and glutaminase (GLS). Taken together, the current study has successfully established an animal model of Bungarus multicinctus bite, and further identified the links between the metabolic perturbations and the pathophysiology and the potential diagnostic biomakers of Bungarus multicinctus bite, which provided valuable insights for studying the mechanism of Bungarus multicinctus bite.

Bungarus andamanensis Biswas and Sanyal, 1978 (Fig. 8A & B, 12C) Material studied: Four adults, CIARI uncatalogued 1–4, from Sippighat, South Andaman (Fig.4G). Morphology: Dorsal scales smooth, in 15:15:15 rows; vertebral scales hexagonal and enlarged; ventrals 188- 196; subcaudals 42–45, undivided; anal single; supralabials 7, 3–4 entering orbit; infralabials 7, 1–4 touching genials; preocular 1; postoculars 2; loreal absent; temporals 1+2. Dorsum lustrous black with 36–45 pale yellow to creamy white bands all over the body and 8–12 bands on the tail. Hemipenis everted in one specimen; organ short, 6 mm long, of conical shape, longer than broad and not lobed. Distal end much broader than proximal end, with intense flounces towards the tip pointing posteriorly. Sulcus spermaticus visible as a groove ventrally. Distribution: An endemic species. Several individuals were encountered in a variety of habitats ranging from evergreen forests, secondary forests, and mangrove swamps to human habitations on South Andaman, Havelock, Long Island, Little Andaman, Middle and North Andaman Islands. Often seen near human habitation close to forests and has been seen crossing roads at night. One gravid female was recorded in December. Status: Common., Published as part of Chandramouli, S. R., 2022, Snake fauna of the Andaman Islands, Bay of Bengal-A review of species richness taxonomy, distribution, natural history and conservation status, pp. 301-331 in Zootaxa 5209 (3) on page 312, DOI: 10.11646/zootaxa.5209.3.1, http://zenodo.org/record/7329609, {"references":["Biswas, S. & Sanyal, D. P. (1978) A new species of krait of the genus Bungarus Daudin, 1803 (Serpents: Elapidae) from the Andaman Island. Journal of the Bombay Natural History Society 75 (1), 179 - 183."]}

Objectives: The present study focuses on development of novel Metal-Herbal Nano composite (MHNC) formulation against Naja naja and Bungarus Caeruleus venoms on animal models. Methods: The venoms were procured from Irula snake catchers society in lyophilised forms. Herbal-metal nanocomposites were prepared in 1:1 combinations. Single strength concentration of Leucas zeylanica extracts and copper oxide metal nanoparticles was used to attain these ratio’s. In vivo venom neutralization analysis was carried out in Swiss albino mice. The LD50 and ED50 of the MHNC were determined. Results: The LD50 of the Naja naja venom was observed to be 0.19μg/g and 0.174μg/g for Bungarus Caeruleus venom. The ED50 of the MHNC against Naja naja venom was observed to be 14.22mg and 63.39mg for Bungarus Caeruleus venom. Conclusion: The MHNC developed in this study has significant venom neutralization potential against Naja naja and Bungarus Caeruleus venoms. Therefore, the MHNC can be used for development of anti-venom drugs.

Background Venomous snake bites cause acute medical emergencies and are fatal. India accounts for large proportion of snake-bite deaths globally. Medically important ‘BIG FOUR’ snakes of India are Bungarus caeruleus (krait), Naja naja (cobra), Echis carinatus (saw-scaled viper) and Daboia russelii (Russell's viper). Polyherbal formulations have been proved to be effective in treatment of diseases than a single formulation. Objective(s) To evaluate aqueous ethanolic extract cocktail of Azadirachata indica, Butea monosperma, Citrus limon, Clerodendrum serratum and Areca catechu for antidote potential against BIG FOUR venoms in ex vivo and in vivo model. Materials and methods Anti-hemorrhagic and venom neutralization studies were performed in seven-day old chick embryo model for ex vivo studies. In vivo studies were performed using male Swiss albino mice for antivenom potential of herbal cocktail by performing anti-edematic, anti-hemorrhagic, anti-myotoxic activity, and venom neutralization. Results Herbal cocktail exhibited differential venom inhibition potential against four venoms tested. Hemorrhagic activity was completely neutralized by the herbal cocktail; myotoxic activities of krait and Russell's viper venom were neutralized; while anti-edematic activity was observed for krait and cobra venom. Herbal cocktail completely neutralized venom lethality (3∗LD50) of krait and saw-scaled viper venom. Conclusion Inhibitions of various venom components of all four venoms suggests presence of phytochemicals in herbal cocktail with therapeutic properties. Further studies would help in the development of a formulation as a first-aid towards treatment of snake bite victims.

Snakebite envenoming is a serious and life-threatening but neglected problem in the tropics. The focus in the Indian subcontinent is usually on the Indian cobra (Naja naja), common krait (Bungarus caeruleus), Russell's viper (Daboia russelii), and Indian saw-scaled viper (Echis carinatus). The Indian polyvalent antivenom contains hyperimmunized horse antibodies against only these 4 species. However, regional intraspecific variations are important in viper envenomings, leading to marked differences in clinical presentation and response to the available polyvalent antivenom. Echis carinatus sochureki, a subspecies of Echis carinatus, has been linked to serious morbidity in the Thar Desert regions of Rajasthan, although consistent reports are lacking. We report a patient with prolonged venom-induced consumption coagulopathy owing to Echis carinatus sochureki envenoming who did not respond to Indian polyvalent antivenom in Jodhpur, India. Features of local and hemotoxic envenoming resolved after a week with supportive care. Echis sochureki venom has been shown to be different from Echis carinatus in terms of composition and in vitro neutralization by antivenom. Clinicians in the tropical desert regions must suspect Echis sochureki envenoming in the setting of nonresponsiveness to Indian polyvalent antivenom. This will help optimize antivenom use in these patients, preventing potentially life-threatening antivenom associated reactions. Because the usefulness of Indian polyvalent antivenom appears to be limited in this setting, there is an urgent need to advocate for region-specific antivenom or monovalent antivenom for this area.

India has recorded the maximum snakebite deaths in the world. Intravenous administration of polyvalent antivenom (PAV) raised against the 'Big Four' venomous snakes of India (Naja naja, Daboia russelli, Echis carinatus, and Bungarus caeraleus) is the only choice of treatment. The WHO has recommended the evaluation of quality and safety of commercial antivenom by in vitro laboratory tests prior to their pre-clinical evaluation in animal model and therapeutic use. Therefore, in this study an attempt has been made to evaluate the quality of commercial polyvalent antivenom produced in India by simple, and affordable laboratory tests. Proteomic analysis revealed that PAVs contained 78.7-94.8% IgG/F(ab')2 and small quantities of plasma proteins. The PAVs showed batch-to-batch variations with varying amounts of undigested IgG and its aggregates, and moderate complement activation. However, absence of IgE, negligible endotoxin contamination, and recommended limit of preservative (cresol) in PAVs were observed. The PAVs contain varying proportions and least amount of venom-specific antibodies against venoms of the 'Big Four' snakes from different locales of India, and against eastern India N. kaouthia venom, respectively. The importance of independent in vitro laboratory tests for the quality control and safety assessment for improving the quality of Indian commercial PAV is reinforced.

Kraits of the genus Bungarus Daudin 1803 are widely known venomous snakes distributed from Iran to China and Indonesia. Here, we use a combination of mitochondrial DNA sequence data and morphological data to describe a new species from Yingjiang County, Yunnan Province, China: Bungarus suzhenaesp. nov. Phylogenetically, this species forms a monophyletic lineage sister to the Bungarus candidus/multicinctus/wanghaotingi complex based on cyt b and ND4 genes but forms a sister species pair with the species B. magnimaculatus Wall & Evans, 1901 based on COI gene fragments. Morphologically, B. suzhenaesp. nov. is similar to the B. candidus/multicinctus/wanghaotingi complex but differs from these taxa by a combination of dental morphology, squamation, coloration pattern, as well as hemipenial morphology. A detailed description of the cranial osteology of the new species is given based on micro-CT tomography images. We revised the morphological characters of B. candidus/multicinctus/wanghaotingi complex and verified the validity of three species in this complex. The distribution of these species was revised; the records of B. candidus in China should be attributed to B. wanghaotingi. We also provide an updated key to species of Bungarus.

There are an estimated 5.4 million snakebite cases every year. People with snakebite envenoming suffer from severe complications, or even death. Although some review articles cover several topics of snakebite envenoming, a review of the cases regarding cerebral complications, especially rare syndromes, is lacking. Here, we overview 35 cases of snakebite by front-fanged snakes, including Bothrops, Daboia, Cerastes, Deinagkistrodon, Trimeresurus, and Crotalus in the Viperidae family; Bungarus and Naja in the Elapidae family, and Homoroselaps (rare cases) in the Lamprophiidae family. We also review three rare cases of snakebite by rear-fanged snakes, including Oxybelis and Leptodeira in the Colubridae family. In the cases of viper bites, most patients (17/24) were diagnosed with ischemic stroke and intracranial hemorrhage, leading to six deaths. We then discuss the potential underlying molecular mechanisms that cause these complications. In cases of elapid bites, neural, cardiac, and ophthalmic disorders are the main complications. Due to the small amount of venom injection and the inability to deep bite, all the rear-fanged snakebites did not develop any severe complications. To date, antivenom (AV) is the most effective therapy for snakebite envenoming. In the six cases of viper and elapid bites that did not receive AV, three cases (two by viper and one by elapid) resulted in death. This indicates that AV treatment is the key to survival after a venomous snakebite. Lastly, we also discuss several studies of therapeutic agents against snakebite-envenoming-induced complications, which could be potential adjuvants along with AV treatment. This article organizes the diagnosis of hemotoxic and neurotoxic envenoming, which may help ER doctors determine the treatment for unidentified snakebite.

Snake-bite gives rise to significant socio-economic, and limitations of antivenom have raised the necessity for the development of novel neutralising compounds. The present study concentrates on evaluating the neutralisation potential of Copper nanoparticles, Leucas zeylanica leaf extracts and Metal-Herbal Nanocomposite (MHNC) against Naja naja and Bungarus caeruleus venoms. The antigenic potency of snake venoms was determined by gel precipitation test. Leucas zeylanica leaves were used for extraction, and extracts were further purified by column chromatography. Size of Leucas zeylanica extract was found to be 86.7±7.1nm and 249.4±27.3nm for MHNC. Zeta potential values determined the Stability of the MHNC. MHNC showed higher neutralisation to venoms than the Leucas zeylanica extracts. In direct hemolysis, the efficiency of Naja naja and Bungarus caeruleus venoms treated with Leucas zeylanica leaf extract was found to be 30% and 27.2%, whereas MHNC showed 16.1% against Naja naja venom and 17.1% Bungarus caeruleus venom. From Indirect analysis, Naja naja and Bungarus caeruleus venom were able to produce 28mm and 26mm diameter hemolytic halo zones. On treatment with MHNC, the zones were reduced to 15mm and 11mm than Leucas zeylanica extract. This result indicates the MHNC can be used for the treatment of snake bites.

Objective We aimed to investigate the effects of snake-derived phospholipase A2 inhibitor (PLA2) from Sinonatrix percarinata and Bungarus multicinctus on acute pancreatitis in vivo and in vitro and assess the mechanisms. Methods The levels of platelet-activating factor (PAF) and tumor necrosis factor (TNF)-α were detected by ELISA, and the characteristics of autophagy were detected by transmission electron microscopy and Western blotting (LC3, p62, and ATG5). Results In vitro experiments showed that PLA2 treatment caused obvious formation of autophagic bodies. By contrast, Sinonatrix and Bungarus peptides reduced the number of autophagic bodies. The concentrations of PAF and TNF-α, and the expressions of p62, autophagy-related 5 (ATG5), and microtubule-associated protein 1A/1B-light chain 3 (LC3)II/LC3I in the PLA2-treated group were significantly higher than in the control group (P

Background: Deinagkistrodon acutus (D. acutus) and Bungarus multicinctus (B. multicinctus) as traditional medicines have been used for hundreds of years in China. The venoms of these two species have strong toxicity on the victims. Objective: The objective of this study is to reveal the profile of venom proteins and peptides of D. acutus and B. multicinctus. Method: Ultrafiltration, SDS-PAGE coupled with in-gel tryptic digestion and Liquid Chromatography- Electrospray Ionization-Tandem Mass Spectrometry (LC-ESI-MS/MS) were used to characterize proteins and peptides of venoms of D. acutus and B. multicinctus. Results: In the D. acutus venom, 67 proteins (16 protein families) were identified, and snake venom metalloproteinases (SVMPs, 38.0%) and snake venom C-type lectins (snaclecs, 36.7%) were dominated proteins. In the B. multicinctus venom, 47 proteins (15 protein families) were identified, and three-finger toxins (3FTxs, 36.3%) and Kunitz-type Serine Protease Inhibitors (KSPIs, 32.8%) were major components. In addition, both venoms contained small amounts of other proteins, such as Snake Venom Serine Proteinases (SVSPs), Phospholipases A2 (PLA2s), Cysteine-Rich Secreted Proteins (CRISPs), 5'nucleotidases (5'NUCs), Phospholipases B (PLBs), Phosphodiesterases (PDEs), Phospholipase A2 Inhibitors (PLIs), Dipeptidyl Peptidases IV (DPP IVs), L-amino Acid Oxidases (LAAOs) and Angiotensin-Converting Enzymes (ACEs). Each venom also had its unique proteins, Nerve Growth Factors (NGFs) and Hyaluronidases (HYs) in D. acutus, and Cobra Venom Factors (CVFs) in B. multicinctus. In the peptidomics, 1543 and 250 peptides were identified in the venoms of D. acutus and B. multicinctus, respectively. Some peptides showed high similarity with neuropeptides, ACE inhibitory peptides, Bradykinin- Potentiating Peptides (BPPs), LAAOs and movement related peptides. Conclusion: Characterization of venom proteins and peptides of D. acutus and B. multicinctus will be helpful for the treatment of envenomation and drug discovery.

Identification of snake venoms is a vital step in the treatment of fatal snakebites. In this study, we use the gold-thiolate interaction between a cysteine residue and gold nanoparticles to establish a SERS method for the differentiation of the venoms of Trimeresurus stejnegeri and Bungarus multicinctus. We confirm the preference of gold nanoparticles over silver for the SERS study of snake venoms by a binding experiment that also functions to differentiate the two venom samples by colorimetry and UV-vis spectroscopy. We report the SERS spectra of Trimeresurus stejnegeri and Bungarus multicinctus venoms for the first time. The spectra display distinct SERS signatures of the snake venoms on bone-shaped gold nanoparticles made with a house recipe. These signatures correlate to selected segments of the venom proteins due to the anchoring effect of the gold-cysteine bond. The method is quick as it accomplishes in situ isolation of the structure of interest to avoid tedious purification of the samples. The location of the interactive cysteine residue makes a novel characteristic of proteins in general.

Bungarus multicinctus is one of the top ten venomous snakes in China. Its venom is mainly neurotoxin-based. Novel antivenom drugs need to be further researched and developed.This study aimed to explore the molecular mechanism of Cynanchum paniculatum in treating Bungarus multicinctus bites based on network pharmacology.Seven potential active components (cynapanoside C, cynatratoside B, tomentolide A, sitosterol, sarcostin, tomentogenin, and paeonol) and 286 drug targets were obtained, including 30 key targets for the treatment of bungarotoxin toxicity. The active components mainly acted on PIK3CA, MAPK1, MAP2K1, JAK2, FYN, ACHE, CHRNA7, CHRNA4, and CHRNB2, and they antagonized the inhibitory effect of bungarotoxin on the nervous system through cholinergic synapses and the neurotrophin signaling pathway.Cynanchum paniculatum exerts a therapeutic effect on Bungarus multicinctus bites through multiple active components, multiple targets, and multiple pathways. The findings provide a theoretical basis for the extraction of active components of Cynanchum paniculatum and for related antivenom experiments.

Sunagar et al. (2021: 1) erected the new snake species Bungarus romulusi on the basis of the molecular study of mitochondrial markers, venom proteomics and venom gland transcriptomics. Unfortunately, this new specific nomen is nomenclaturally unavailable, and should therefore not be used as valid as long as it has not been properly published.

South Asian subcontinent is among the highest burden areas in respect of snake bite. India is commonest place in this subcontinent with mortality following snake envenomation. The four most important venomous snakes in India are cobra (Naja naja) and common krait (Bungarus caeruleus), which are neurotoxic, and the saw-scaled viper (Echis carinatus) and Russell’s viper (Daboia russelii), which are hemotoxic1. Various autonomic dysfunction following neuro-paralytic envenomation with krait and cobra has been reported. Timely recognition and appropriate management of snake envenomation along with autonomic dysfunction helps to reduce morbidity and mortality. Keywords: Autonomic dysfunction, hypertension, antisnake venom, blood pressure.

Snakebites are common in India and the most common neurotoxic snakebites in India are due to Common krait (Bungarus caeruleus) and cobra (Naja naja). Severe envenomation may mimic brain death or a locked-in state with flaccid paralysis in a descending manner and total ophthalmoplegia. Usually, patients who receive timely antivenom and ventilator support recover completely without any sequalae. We are reporting two cases of krait bite with an unusually long period of flaccid paralysis, which required prolong ventilation. While case 1 required 10 days of mechanical ventilation followed by 5 days of non-invasive ventilation, case 2 required 11 days of mechanical ventilation followed by 5 days of non-invasive ventilation. Both the cases had delayed recovery and residual weakness at 3-month follow up. These case reports suggest that krait bite may cause prolong neuromuscular weakness in children, which has implications for both acute and chronic management.

To search for compounds with antiprotozoal activity, effects of snake venoms on the ciliates Tetrahymena pyriformis was studied. T. pyriformis from subkingdom of Protozoa, including the protozoal pathogens, was used as a model organism to select the venoms that are the most active against parasitic protozoans. Various concentrations of venoms were added to the cells, and the cells that survived after 24 h were counted. Among the six snake species from the Viperidae family, the venom of the viper Vipera berus, which completely killed the cells at 49 μg/mL, was the most active. Among four species from the Elapidae family, the previously studied cobra venoms containing cytotoxins with strong antiprotozoal activity as well as the venom of krait Bungarus multicinctus (10 μg/mL) were the most active. The venoms of the pit vipers and Nikolsky's viper did not show any activity at 12.5 mg/mL. Thus, the venoms of V. berus and B. multicinctus are promising for the isolation of new antiprotozoal compounds.

Snake envenomation is a severe problem in Sri Lanka (SL) and Indian polyvalent antivenom (PAV) is mostly used for treating snakebite albeit due to geographical variation in venom composition, Indian PAV shows poor efficacy in neutralizing the lethality and toxicity of venom from the same species of snakes in SL. Therefore, the quality and in vivo venom neutralization potency of a country-specific PAV produced against the venom of the five most medically important snakes of SL (Daboia russelii, Echis carinatus, Hypnale hypnale, Naja naja, Bungarus caeruleus) was assessed. LC-MS/MS analysis of two batches of PAV showed the presence of 88.7–97.2% IgG and traces of other plasma proteins. The tested PAVs contained minor amounts of undigested IgG and F(ab′)2 aggregates, showed complement activation, were devoid of IgE, endotoxin, and content of preservative was below the threshold level. Immunological cross-reactivity and in vitro neutralization of enzymatic activities, pharmacological properties demonstrated superior efficacy of SL PAV compared to Indian PAV against SL snake venoms. The in vivo neutralization study showed that the tested PAVs are potent to neutralize the lethality and venom-induced toxicity of SL snake venoms. Therefore, our study suggests that introduction of SL-specific PAV will improve snakebite management in SL.

Animal movement can impact human-wildlife conflict through a variety of features: increased movement can lead to greater chance of encounter, remaining still can lead to greater or lesser detection, and activity can modulate their impact on humans. Here we assess the movement and space use of the highly venomous and medically important Malayan krait (Bungarus candidus) on a suburban university campus. We radio- tracked 14 kraits for an average of 114 days (min: 19, max: 218), with an average of 106 fixes (min: 21, max: 229). We assessed movement pathways and activity with dynamic Brownian Bridge Movement models, evaluated patterns of revisitation to identify site fidelity, and determined whether individuals selected for particular land-use types with Integrated Step Selection Functions. Most individuals displayed some level of attraction to buildings (n = 10) and natural areas (n = 12); we identified a similar unambiguous pattern of attraction to buildings and natural areas at the population level (of our sample). Snakes remained in shelter sites for long periods of time (max = 94 days) and revisited sites on average every 15.45 days. Over 50% of fixes were within human settlements and 37.1% were associated with buildings. We found generally seasonal patterns of activity, with higher activity in wet seasons (when classes are typically in session on campus), and lower activity in the hot season (when there are a number of short breaks causing students to leave campus). These results show frequent proximity between Malayan kraits and humans at Suranaree University of Technology, Thailand; thereby, suggesting a near constant potential for human- wildlife conflict. Despite the fact that no snakebites from this species occurred at the university during our study period, substantial education and awareness training should be considered to raise awareness to ensure continued coexistence on campus.

An understanding of the spatial ecology of species living in and around human-dominated habitats is needed to develop conflict mitigation strategies and predict how organisms cope with ongoing anthropogenic habitat alteration. Here we present the results of a six-month telemetry study in Thailand of the banded krait (Bungarus fasciatus), a venomous elapid snake. We quantified home range size and habitat use of three adult kraits (1 male, 2 females) in an agricultural habitat. The kraits travelled an average of 47.20 m ± 23.54 m between shelter sites and occupied home ranges of on average 21.45 ha ± 19.56 ha (95 % dynamic Brownian Bridge Movement Models). They preferred to seek day time refuge amongst less disturbed habitat features, including waterways and rice paddy dykes, while avoiding routinely disturbed monoculture fields. The two female kraits also exhibited reduced movement while simultaneously attending their nests in a shared burrow complex. Fifteen neonates were observed emerging from the nest site. We recommend further investigation of banded krait reproductive ecology and ability to persist in heavily disturbed habitats.

Background: Snakes are found on every continent in the world except Antarctica, and on smaller land masses. Being ecologically important, they also cause a large number of bites, leading to millions of deaths. Mitochondrial and nuclear gene sequences are being used to identify, characterize, and infer genetic biodiversity among different snake species. Furthermore, phylogenetics helps in inferring the relationships and evolutionary histories among these species. Black cobra is one of the four most venomous snakes in Pakistan. Four mitochondrial (ND4, Cytochrome b, 12S rRNA, and 16S rRNA) and four nuclear (C-mos, RAG-1, BDNF, and NT3) genes were used to trace diversity and infer the phylogenetic relationship of black cobra in Pakistan. Results: Almost similar phylogenies were obtained through maximum likelihood and Bayesian inference, showing two species of cobra in Pakistan, namely, black cobra (Naja naja) and brown cobra (Naja oxiana). All Naja species were divided into three clades: black cobra (N. naja) and brown cobra (N. oxiana) cladding with different species of Naja; N. naja (Pakistan) cladding with N. naja from Nepal; and N. oxiana showed close relationship with Naja kaouthia from Thailand and Naja siamensis from Thailand. Conclusion: It was confirmed genetically that there are two cobra species in Pakistan, i.e., black and brown cobras. This study will help in not only genetic conservation but also developing anti-venom against snake species.How to cite: Ashraf MR, Nadeem A, Smith EN, et al. Molecular phylogenetics of Black Cobra (Naja naja) in Pakistan. Electron J Biotechnol 2019;42. https://doi.org/10.1016/j.ejbt.2019.10.005. Keywords: Bungarus, Cobra, Mitochondrial DNA, Naja, Nuclear DNA, Pakistan, PCR, Phylogenetics, Reptiles, Snakes, Venomous snakes

Snake bite envenoming is a worldwide problem which affects the health and safety of the public. The classical antivenom methods are difficult to be used immediately and accurately. Based on this situation, a new metal-organic framework and hydrogel based first-aid treatment was first set up in this study as a facile, economic, effective, and universal alternative strategy. According to the subsequent in vitro and in vivo experiments, this dual effective (block and absorption) strategy was proven to be useful in dealing with three different representative snake venoms (Bungarus fasciatus, Protobothrops mucrosquamatus, and cobra venom).

India is the home to several venomous snake species which are responsible for the highest number of snake envenomings and deaths in the world. Nevertheless, the ‘Big Four’ species of snakes namely Russell’s viper (Daboia russelii), spectacled cobra (Naja naja), common krait (Bungarus caeruleus), and saw-scaled viper (Echis carinatus) account for most cases of envenoming, morbidity, and mortality primarily in the rural parts of the country. The composition of these snake venoms have been determined using conventional biochemical approaches. However, these studies faced the inevitable limitation of identification of non-enzymatic and low abundance venom toxins. In the last 5 years, mass spectrometry investigations of Indian snake venoms have provided comprehensive information on their venom proteomes. In addition, proteomic analysis has also been employed to identify and characterize snake venom protein complexes and serves as a tool for screening of candidate pharmacologically active drug prototypes from snake venoms. Further, mass spectrometry analysis has also found its way for the identification and quantification of poorly immunogenic venom toxins against commercial antivenoms and this information is of paramount importance for improved design and production of snake antivenoms for better hospital management of snakebite victims. In this review article, we have focused the recent advances in snake venom proteomics research in India that has facilitated the understanding of the pathophysiology of snake envenomation, provided insights for improvement for antivenom production and also discover drug prototypes from snake venom.

Objective: Snake envenomation is a major cause of death and disability in the developing countries. In India and neighboring countries, the four venomous snakes of concern include – Indian cobra(Naja naja), Common Krait (Bungarus caeruleus); Russell's Viper (Daboia russelli); Saw Scaled Viper (Echis carinatus). We describe the management protocol for snakebite treatment in a tertiary care hospital of Paschim Medinipur district, West Bengal based on case reports of subjects admitted and treated in Ghatal Subdivisional Hospital(GSH) during 2013–2016. Methods &materials: We developed a structured data collection form to record demographic and clinical details of patients hospitalized at GSH between 1 January 2013 through 31 December 2016. Results: Snake bite cases in Ghatal Subdivisional hospital(GSH) were assessed during the period 2013–2016. A total 18 deaths due to snakebite has been reported from this tertiary care hospital during the period. Total patients admitted in this hospital with snakebite is 1160 during the period 2013–2016. In 80% of the cases the lower extremities were affected. Preliminary first aid was provided in 45% cases. About 65% of the affected victims suffered snakebite in the morning hours. Some of the recommended drugs that were prescribed by the physicians of GSH were neostigmine, atropine, adrenaline, hydrocortisone, Amoxicillin. WBCT20 and Urea, Creatinine level were routinely performed. Conclusion: Hospital studies are a key source of information about snake bites. The ready availability and appropriate use of AVS, close monitoring of patients, the institution of ventilator support and if required, early referral to a larger hospital all help to reduce the mortality. Thus knowledge of the varied clinical manifestations of snake bite is important for effective management in hospitals by a complete health care team. Keywords: Snake envenomation, Russell's viper

The Red-headed Krait (Bungarus flaviceps) is a medically important venomous snake species in Southeast Asia, while there is no specific antivenom available for its envenoming. This study investigated the venom composition through a decomplexation proteomic approach, and examined the immunoreactivity as well as cross-neutralization efficacy of two hetero-specific krait antivenoms, Bungarus candidus Monovalent Antivenom (BcMAV) and Bungarus fasciatus Monovalent Antivenom (BfMAV), against the venom of B. flaviceps from Peninsular Malaysia. A total of 43 non-redundant proteoforms belonging to 10 toxin families were identified in the venom proteome, which is dominated by phospholipases A

Bungarus multicinctus is a widely distributed and medically important elapid snake that produces lethal neurotoxic venom. To study and enhance existing antivenom, we explore the complete repertoire of its toxin genes based on de novo chromosome-level assembly and multi-tissue transcriptome data. Comparative genomic analyses suggest that the three-finger toxin family (3FTX) may evolve through the neofunctionalization of flanking LY6E. A long-neglected 3FTX subfamily (i.e., MKA-3FTX) is also investigated. Only one MKA-3FTX gene, which evolves a different protein conformation, is under positive selection and actively transcribed in the venom gland, functioning as a major toxin effector together with MKT-3FTX subfamily homologs. Furthermore, this lethal snake may acquire self-resistance to its β-bungarotoxin via amino acid replacements on fast-evolving KCNA2. This study provides valuable resources for further evolutionary and structure-function studies of snake toxins, which are fundamental for the development of effective antivenoms and drug candidates.

Bungarus candidus (Linnaeus, 1758) Figure 6D Materials examined. MALAYSIA – Kedah • Batu Hampar Recreational Forest; 05.1966°N, 100.5827°E; 40 m a.s.l.; 23.IX.2018; Evan S. H. Quah, Hong Zijia leg.; 1 &male;, USMHC 2453 • same locality; 19.X.2018; Evan S. H. Quah, Hong Zijia leg.; 1 &female;, USMHC 2483. Identification and natural history. One adult male (USMHC 2453, SVL 790 mm, TL 110 mm) and one adult female (USMHC 2483, SVL 792 mm, TL 70 mm) matched Tweedie’s (1983) and Das’s (2015) diagnosis in having a robust body; head indistinct from neck; single preocular; two postoculars; loreal absent; seven supralabials, where the 3rd and 4th touches the eye; seven infralabials; small eye; rounded pupil; short tail; smooth dorsals; 15 midbody scale rows; 194–237 ventrals; 37– 56 single subcaudals; anal scale entire; black dorsum with broad white cross bars on body and tail, where the black interspaces are much wider than the white bands anteriorly, slightly wider or equal posteriorly and on the tail; white bands scattered with black spots; supralabials and venter white and immaculate and dark brown spots on subcaudals. USMHC 2453 was found crawling along puddle by trail while USMHC 2483 was found in the leaf litter along trail, eating a skink, Eutropis multifasciata (USMHC 2484) at night., Published as part of Zijia, Hong, Anuar, Shahrul, Grismer, L. Lee & Quah, Evan S. H., 2021, Preliminary report on the herpetofaunal diversity of Batu Hampar Recreational Forest, Kedah, Malaysia, pp. 791-814 in Check List 17 (3) on page 803, DOI: 10.15560/17.3.791, {"references":["Linnaeus C (1758) Systema naturae per regna tria naturae, secundum classes, ordines, genera, species, cum characteribus, differentiis, synonymis, locis (Vol. 1). 10 th edition. Laurentii Salvii, Stockholm, Sweden, 824 pp.","Tweedie MWF (1983) The snakes of Malaya. Singapore National Printers, Singapore, 167 pp."]}

In the Southern Asian countries, snakebite takes a substantial toll in terms of human life, inflicts acute morbidity and long term disability both physical and psychological, and therefore represents a neglected socio-economic problem and severe health issue that requires immediate medical attention. The 'Big Four' venomous snakes, viz. Daboia russelii, Naja naja, Bungarus caeruleus and Echis carinatus, are prominent, medically important species and are the most dangerous snakes of this region; therefore, the commercial polyvalent antivenom (PAV) contains antibodies against the venoms of these snakes. However, envenomations by species other than the 'Big Four' snakes are grossly neglected, and PAV is only partially effective in neutralizing the venom of these snakes. Many issues confounding effective treatment of snakebite are discussed in this review, and these hurdles preventing successful treatment of snakebite must be addressed. However, in South Asian countries, the pre-hospital treatment and appropriate first aid are equally important to mitigate the problem of snakebite and therefore, these issues are also highlighted here. Further, this review suggests a roadmap and guidelines for the prevention of snakebite and improvement of hospital management of snakebite in these Southern Asian countries.

Complex target SELEX always have been an intriguing approach to the scientific community, as it offers the potential discovery of novel biomarkers. We herein successfully performed SELEX on Bungarus caeruleus venom to develop a panel of highly affine aptamers that specifically recognizes the B. caeruleus (common krait) venom and was able to discriminate the B. caeruleus venom from Cobra, Russell's, and Saw-scaled viper's venom. The aptamers generated against the crude venom also lead to the identification of the specific component of the venom, which is β-Bungarotoxin, a toxin uniquely present in the B. caeruleus venom. The best performing aptamer candidates were used as a molecular recognition element in a paper-based device and were able to detect as low as 2 ng krait venom in human serum background. The developed aptamer-based paper device can be used for potential point-of-care venom detection applications due to its simplicity and affordability.

Snakebite envenoming is a global public health problem. The pediatric population poisoned by snakebite envenoming has different features than adult patients. Bungarus multicinctus is a highly venomous species of the elapid snake. The documented clinical presentations following Bungarus multicinctus envenoming are minimal local reactions, respiratory failure, general pain, and life-threatening hyponatremia. We present an uncommon case of Bungarus multicinctus envenomation in a girl with unusual clinical findings, including severe tissue necrosis and thrombocytopenia with coagulopathy.

Ceylon krait (Bungarus ceylonicus) of the family Elapidae is a highly venomous endemic species inhabiting in the wet zone and some parts of the intermediate climatic zones of Sri Lanka. Clinical records of its bites are rare and limited to five case reports in the literature. It is of interest to note that there are several non-venomous snakes in Sri Lanka having similar morphological appearance to kraits causing identification difficulties which lead to unnecessary and unindicated administration of antivenom. We report two paediatric cases of proven Ceylon krait bites and three adult patients with similar looking non-venomous snakebites. These children were 1½ and 13 years old and developed neuroparalysis without progressing to respiratory failure and recovered. Both the children were administered Indian polyvalent antivenom which has not developed against endemic Ceylon krait venom. The two adult patients also received antivenom due to the misidentification without clinical and laboratory evidence of envenoming.

Bungarus multicinctus, the Chinese krait, is a highly venomous elapid snake which causes considerable morbidity and mortality in southern China. B. multicinctus venom contains pre-synaptic PLA2 neurotoxins (i.e., &beta, bungarotoxins) and post-synaptic neurotoxins (i.e., &alpha, bungarotoxins). We examined the in vitro neurotoxicity of B. multicinctus venom, and the efficacy of specific monovalent Chinese B. multicinctus antivenom, and Australian polyvalent elapid snake antivenom, against venom-induced neurotoxicity. B. multicinctus venom (1&ndash, 10 &mu, g/mL) abolished indirect twitches in the chick biventer cervicis nerve-muscle preparation as well as attenuating contractile responses to exogenous ACh and CCh, but not KCl. This indicates a post-synaptic neurotoxic action but myotoxicity was not evident. Given that post-synaptic &alpha, neurotoxins have a more rapid onset than pre-synaptic neurotoxins, the activity of the latter in the whole venom will be masked. The prior addition of Chinese B. multicinctus antivenom (12 U/mL) or Australian polyvalent snake antivenom (15 U/mL), markedly attenuated the neurotoxic actions of B. multicinctus venom (3 &mu, g/mL) and prevented the inhibition of contractile responses to ACh and CCh. The addition of B. multicinctus antivenom (60 U/mL), or Australian polyvalent snake antivenom (50 U/mL), at the t90 time point after the addition of B. multicinctus venom (3 &mu, g/mL), did not restore the twitch height over 180 min. The earlier addition of B. multicinctus antivenom (60 U/mL), at the t20 or t50 time points, also failed to prevent the neurotoxic effects of the venom but did delay the time to abolish twitches based on a comparison of t90 values. Repeated washing of the preparation with physiological salt solution, commencing at the t20 time point, failed to reverse the neurotoxic effects of venom or delay the time to abolish twitches. This study showed that B. multicinctus venom displays marked in vitro neurotoxicity in a skeletal muscle preparation which is not reversed by antivenom. This does not appear to be related to antivenom efficacy, but due to the irreversible/pseudo-irreversible nature of the neurotoxins.

The Indian common krait (Bungarus caeruleus) or blue krait is one of the members of the “Big Four” venomous snakes of India. Black or bluish-black in color, it has an average length of 1–1.2 m and possesses a flat, blunt, short head with small eyes, with a neck that is barely visible. The snake has a distinctive feature of yellow and black cross-bands along its backbone. The common krait is distributed in the Indian subcontinent, from Afghanistan, the Sindh province of Pakistan, to the plains of West Bengal in eastern India, Bangladesh, and Nepal, though another species of krait (Bungarus fasciatus, the banded krait) predominates in the northeastern part of India. This snake is nocturnal (night dweller) and therefore the maximum number of krait bites are reported during late night. Although the venom is very potent, because of the poor yields, the biochemical composition of Indian common krait venom has not been analyzed in great detail. Only a few toxins from B. caeruleus venom have been purified and characterized though recent proteomic analyses have provided more comprehensive data on the occurrence of different toxins in its venom. The venom contains 57 distinct proteins distributed in 12 snake venom toxin families. The proteomic analyses indicate that PLA2 (37.6%) and three-finger toxins (48.3%) are the most abundant enzymatic and nonenzymatic protein families, respectively. A comparison of two independent studies on the proteomic analyses of krait venom from two distinct geographical locales (southern India and Sri Lanka) has shown the differences in venom composition for B. caeruleus in these countries. Similar to Indian cobra bite, Indian krait bite patients also have neuroparalytic symptoms and also frequently experience abdominal pain and cramps due to internal bleeding. Further studies are required to characterize and compare the venom composition of the Indian common krait from different regions.

The evolution of snakes from an ancestral burrowing lizard remains disputed because the missing link between snakes and lizards is yet to be recognized. The available fossil record has led to the postulation that the ancestors of snakes made their first appearance during the Cretaceous Period. In recent decades, several studies have shown the mechanism of limb loss in snakes, though the concept is widely debated and the subject of speculation among evolutionary biologists. About 23–65 million years ago during two-thirds of the Tertiary Period, smaller python-like snakes were most prevalent on earth. Phylogenetic studies have indicated that the evolution of snakes involved a steady trend toward greater surface activity, enlargement of the body size, and an enlarged gape. The rear-fanged colubrid snakes evolved 35 to 55 million years ago; however, the origin of the Viperidae and Elapidae from the colubrid snakes is less clear. One group of researchers proposed a common single origin of the Viperidae and the Elapidae from colubrids while another group claimed that the elapids arose from the opisthoglyphous snakes and the viperids were derived from the proteroglyphous colubrids. In this chapter, different theories from evolutionary biologists are examined to explain the origin and evolution of snakes. The “Big Four” venomous snakes of India (N. naja and B. caeruleus of the Elapidae family, and D. russelii and E. carinatus of the Viperidae family) belong to the Infraorder Caenophidia and they all are members of the front-fanged advanced snakes. In addition, the occurrence of different species of the “Big Four” venomous snakes and their geographical distribution in the Indian subcontinent are discussed.

Bungarus multicinctus, the Chinese krait, is a highly venomous elapid snake which causes considerable morbidity and mortality in southern China. B. multicinctus venom contains pre-synaptic PLA2 neurotoxins (i.e., β-bungarotoxins) and post-synaptic neurotoxins (i.e., α-bungarotoxins). We examined the in vitro neurotoxicity of B. multicinctus venom, and the efficacy of specific monovalent Chinese B. multicinctus antivenom, and Australian polyvalent elapid snake antivenom, against venom-induced neurotoxicity. B. multicinctus venom (1–10 μg/mL) abolished indirect twitches in the chick biventer cervicis nerve-muscle preparation as well as attenuating contractile responses to exogenous ACh and CCh, but not KCl. This indicates a post-synaptic neurotoxic action but myotoxicity was not evident. Given that post-synaptic α-neurotoxins have a more rapid onset than pre-synaptic neurotoxins, the activity of the latter in the whole venom will be masked. The prior addition of Chinese B. multicinctus antivenom (12 U/mL) or Australian polyvalent snake antivenom (15 U/mL), markedly attenuated the neurotoxic actions of B. multicinctus venom (3 μg/mL) and prevented the inhibition of contractile responses to ACh and CCh. The addition of B. multicinctus antivenom (60 U/mL), or Australian polyvalent snake antivenom (50 U/mL), at the t90 time point after the addition of B. multicinctus venom (3 μg/mL), did not restore the twitch height over 180 min. The earlier addition of B. multicinctus antivenom (60 U/mL), at the t20 or t50 time points, also failed to prevent the neurotoxic effects of the venom but did delay the time to abolish twitches based on a comparison of t90 values. Repeated washing of the preparation with physiological salt solution, commencing at the t20 time point, failed to reverse the neurotoxic effects of venom or delay the time to abolish twitches. This study showed that B. multicinctus venom displays marked in vitro neurotoxicity in a skeletal muscle preparation which is not reversed by antivenom. This does not appear to be related to antivenom efficacy, but due to the irreversible/pseudo-irreversible nature of the neurotoxins.

Bungarus multicinctus is the most venomous snake distributed in China and neighboring countries of Myanmar, Laos, north Vietnam and Thailand. The high mortality rate of B. multicinctus envenomation is attributed to the lethal components of α-, β-, γ- and κ- bungarotoxins contained in the venom. Although anti-B. multicinctus sera were produced in Shanghai, Taiwan and Vietnam, the most widely clinic used product was term as B. multicinctus antivenin and manufactured by Shanghai Serum Bio-technology Co. Ltd. In the present investigation, high purity α-, β- and γ-bungarotoxins were separately isolated from B. multicinctus crude venom. Rabbit anti- α-, β- and γ-bungarotoxin antisera were prepared by common methods, respectively. LD50 values of α-, β- and γ-bungarotoxins were systematically determined via three administration pathways (intraperitoneal, intramuscular and intravenous injections) in Kunming mice. LD50 values of β-bungarotoxin were closely related with injection routines but those of both α- and γ-bungarotoxins were not dependent on the injection routines. Commercial B. multicinctus antivenin showed strong immunoreaction with high molecular weight fractions of the B. multicinctus but weakly recognized low molecular weight fractions like α- and γ-bungarotoxins. Although B. multicinctus antivenin showed immunoreaction with high molecular weight fractions of Bungarus fasciatus, Naja atra, Ophiophagus hannah venoms but the antivenin only demonstrated animal protection efficacy against O. hannah venom. These results indicated that the high molecular weight fractions of the O. hannah played an important role in venom lethality but those of B. fasciatus and N. atra did not have such a role., Author summary Snakebite envenoming is an important public health problem around the world. Bungarus multicinctus envenomation is regarded as most dangerous animal bitten disease in China and neighboring countries. At present, the most widely clinic used B. multicinctus antisera was term as B. multicinctus antivenin and manufactured by Shanghai Serum Bio-technology Co. Ltd. By using high purity α-, β- and γ-bungarotoxins isolated from B. multicinctus crude venom and prepared corresponding rabbit anti- α-, β- and γ-bungarotoxin antisera, we evaluated the efficacy of B. multicinctus antivenin against the crude venom, isolated bungarotoxins, as well as other 8 medical important Chinese venomous snake venoms. B. multicinctus antivenin showed strong immunoreaction with high molecular weight fractions of the B. multicinctus venom but weakly recognized low molecular weight fractions like α- and γ-bungarotoxins. Moreover, B. multicinctus antivenin can neutralize Ophiophagus hannah venom but not B. fasciatus venom was demonstrated.

Background A comprehensive evaluation of the -omic profiles of venom is important for understanding the potential function and evolution of snake venom. Here, we conducted an integrated multi-omics-analysis to unveil the venom-transcriptomic and venomic profiles in a same group of spine-bellied sea snakes (Hydrophis curtus) from the South China Sea, where the snake is a widespread species and might generate regionally-specific venom potentially harmful to human activities. The capacity of two heterologous antivenoms to immunocapture the H. curtus venom was determined for an in-depth evaluation of their rationality in treatment of H. curtus envenomation. In addition, a phylogenetic analysis by maximum likelihood was used to detect the adaptive molecular evolution of full-length toxin-coding unigenes. Results A total of 90,909,384 pairs of clean reads were generated via Illumina sequencing from a pooled cDNA library of six specimens, and yielding 148,121 unigenes through de novo assembly. Sequence similarity searching harvested 63,845 valid annotations, including 63,789 non-toxin-coding and 56 toxin-coding unigenes belonging to 22 protein families. Three protein families, three-finger toxins (3-FTx), phospholipase A2 (PLA2), and cysteine-rich secretory protein, were detected in the venom proteome. 3-FTx (27.15% in the transcriptome/41.94% in the proteome) and PLA2 (59.71%/49.36%) were identified as the most abundant families in the venom-gland transcriptome and venom proteome. In addition, 24 unigenes from 11 protein families were shown to have experienced positive selection in their evolutionary history, whereas four were relatively conserved throughout evolution. Commercial Naja atra antivenom exhibited a stronger capacity than Bungarus multicinctus antivenom to immunocapture H. curtus venom components, especially short neurotoxins, with the capacity of both antivenoms to immunocapture short neurotoxins being weaker than that for PLA2s. Conclusions Our study clarified the venom-gland transcriptomic and venomic profiles along with the within-group divergence of a H. curtus population from the South China Sea. Adaptive evolution of most venom components driven by natural selection appeared to occur rapidly during evolutionary history. Notably, the utility of commercial N. atra and B. multicinctus antivenoms against H. curtus toxins was not comprehensive; thus, the development of species-specific antivenom is urgently needed.

The 'Big Four' venomous snakes -A comprehensive analysis of the venom proteome composition of the 'Big Four' and other medically important venomous snakes of India and the effect of regional variation in venom composition on immunorecognition and/or neutralization by commercial antivenom was undertaken by searching the literature (from 1985 to date) available in large public databases. Further, mass spectrometric identification of poorly immunogenic toxins of snake venom (against which commercial polyvalent antivenom contains a significantly lower proportion of antibodies) and its impact on antivenom therapy against snakebite are discussed. The application of mass spectrometry to identify protein (toxin) complexes as well as drug prototypes from Indian snake venoms and the clinical importance of such studies are also highlighted.Further detailed clinical and proteomic research is warranted to better understand the effects of regional snake venom composition on the clinical manifestation of envenomation and antivenom therapy and to improve the production of antibodies against poorly immunogenic venom components.

Background Detection and quantification of snake venom in envenomed patients’ blood is important for identifying the species responsible for the bite, determining administration of antivenom, confirming whether sufficient antivenom has been given, detecting recurrence of envenoming, and in forensic investigation. Currently, snake venom detection is not available in clinical practice in Sri Lanka. This study describes the development of enzyme immunoassays (EIA) to differentiate and quantify venoms of Russell’s viper (Daboia russelii), saw-scaled viper (Echis carinatus), common cobra (Naja naja), Indian krait (Bungarus caeruleus), and hump-nosed pit viper (Hypnale hypnale) in the blood of envenomed patients in Sri Lanka. Methodology / Principal findings A double sandwich EIA of high analytical sensitivity was developed using biotin-streptavidin amplification for detection of venom antigens. Detection and quantification of D. russelii, N. naja, B. caeruleus, and H. hypnale venoms in samples from envenomed patients was achieved with the assay. Minimum (less than 5%) cross reactivity was observed between species, except in the case of closely related species of the same genus (i.e., Hypnale). Persistence/ recurrence of venom detection following D. russelii envenoming is also reported, as well as detection of venom in samples collected after antivenom administration. The lack of specific antivenom for Hypnale sp envenoming allowed the detection of venom antigen in circulation up to 24 hours post bite. Conclusion The EIA developed provides a highly sensitive assay to detect and quantify five types of Sri Lankan snake venoms, and should be useful for toxinological research, clinical studies, and forensic diagnosis., Author summary Snakebite is a major medical and public health problem in tropical agricultural world. Detection of the type of snake venom and measurement of venom levels in blood are important for snakebite research, selecting the appropriate antivenom, and assessing venom levels in blood at the clinical setting. Currently, a snake venom detection platform is not available in clinical practice in Sri Lanka. This study aimed to develop a double sandwich enzyme immunoassays (EIA) to differentiate and quantify venoms of Russell’s viper (Daboia russelii), saw-scaled viper (Echis carinatus), common cobra (Naja naja), Indian krait (Bungarus caeruleus), and hump-nosed pit viper (Hypnale hypnale) in blood samples of envenomed patients in Sri Lanka. The EIA developed used biotin-streptavidin amplification for detection of venom antigens and showed high analytical sensitivity. The assay allowed the quantification of venoms of the five species in blood samples from envenomed patients. Low level of cross reactivity was noted between species, except in the case of closely related Hypnale species. The presence of D. russelii venom after antivenom treatment is reported, a finding that has implications in the dosing of antivenom in these envenomings. Lack of specific antivenom for H. hypnale envenoming offered an opportunity of study the remaining venom antigen in circulation up to 24 hr post bite. The EIA developed constitutes a useful tool to detect and quantify the five types of Sri Lankan snake venoms, and should be useful for research purposes, as well as for the diagnosis and therapy evaluation of clinical cases of envenomings in this country, and for forensic purposes.

Bungarus multicinctus, an important traditional Chinese medicine, possesses remarkable medicinal activities, while lots of adulterants from other species were misused as B. multicinctus for its large demand and resource starvation. In order to accurately identify B. multicinctus and its common adulterants such as Sinonatrix annularis, Xenochrophis flavipunctatus, Deinagkistrodon acutus, and Naja atra, a simultaneous identification method was designed with multiplex ligation-dependent probe amplification (MLPA) analysis. Five species-specific MLPA probe-couples for B. multicinctus and its common adulterants were designed based on the universal primer amplified COI sequences, which can specifically detect the five species with no mutual interference, and sensitivity analysis showed as less as 5% B. multicinctus or 8.75% adulterants in the mixed samples can be identified in a MLPA assay, especially, the relative quantity of the adulterants can be also inferred based on the MLPA peak area values. Moreover, the results of the present study confirmed the effectiveness of this technique in terms of simultaneous identification of B. multicinctus and its common adulterants in an assay, which has great potential for ensuring the safety of this commercially valuable snake species.

Venom gland is a highly efficient venom production system to maintain their predatory arsenal. Venom toxins mRNA has been shown to increase abruptly in snake after venom expenditure, while the dynamics of venom accumulation during synthesis are poorly understood. Here, PacBio long-read sequencing, Illumina RNA sequencing (RNA-seq), and label-free proteome quantification were used to investigate the composition landscape and time- and temperature-dependent dynamics changes of the Bungarus multicinctus venom gland system. Transcriptome data (19.5223 Gb) from six adult B. multicinctus tissues were sequenced using PacBio RS II to generate a reference assembly, and average 7.28 Gb of clean RNA-seq data was obtained from venom glands by Illumina sequencing. Differentially expressed genes (DEGs) mainly were protein processing rather than venom toxins. Post-translational modifications provided the evidence of the significantly different proportions of toxins in the venom proteome with the changing of replenishment time and temperature, but constant of venom toxins mRNA in the venom gland transcriptome. Dynamic of toxins and genes involved in venom gland contraction suggesting the formation of the mature venom gland system would take at least 9 days. In addition, 59 toxin processing genes were identified, peptidylprolyl isomerase B of which underwent positive selection in Toxicofera. These results provide a reference for determining the extraction time of venom, production of polyclonal and monoclonal antibody for precise treatment plans of venomous snakebites, and construction of an in vitro synthesis system for snake venom protein.