Comparative Investigation of Five Cobras’ Venoms Analgesic Capacity

Every day, a lot of people suffer from different types and intensity pain. Pain is probably the most common symptomatic reason to seek medical consultation. Unfortunately, despite improved knowledge of underlying mechanisms and better treatments, many people who have any type of pain receive inadequate care and non-effective drugs. Although the pain transmission channels are intensively studied, and the drug market is constantly replenished with new analgesics, it is well known that existing medications for the treatment of pain are often associated with serious side effects and rapid development of tolerance (moderate efficiency, physical dependence, respiratory arrest, suffocation, cardiac arrest, etc.). Thus, there is a need for new, more effective remedies. For this reason, despite the presence of a large number of anti-pain drugs, research and development of more effective and safe means for anaesthesia continue. Natural resources, particularly venoms, are a perspective supplier of antinociceptive and anti-inflammatory medicines. Venoms are complex mixtures of bioactive substances with high selectivity for physiological processes, including modulation of different ion channels, receptors function, and metabolic pathways. Thus, venoms represent an extensive source of molecules for the development of therapeutic agents. The goal of this study was the comparison of antinociceptive effects of five different cobras' venom antinociceptive action of cobras’ venoms was carried out under the same experimental conditions, at the same doses, during acute and inflammatory pain in mice in “formalin test”. To avoid toxic effects, the sublethal dose of each cobra venom (approximately 1/10 LD 50 ) was selected. The behavioural study showed that all tested venoms had a slight sensitizing effect in the acute phase during the first 5 min. In the second, inflammatory phase (16*25min), all tested cobra’s venoms (3μg/0.1ml, intraperitoneal) showed significant antinociceptive action, particularly the Naja naja oxiana venom decreased pain sensitivity by 48.4%, the Naja naja pallida venom by 75.4%, the Naja naja nigricincta venom by 38.5%, the Naja naja kaouthia venom by 33.2%, and the Ophiophagus Hannah venom by 78.3%, (p<0.05). The analgesic capacity of Analgin and Diclofenac under the same conditions were 77.9% and 88.7%, respectively. Thus, the Naja n. pallida and Ophiophagus Hannah venoms have shown the most expressed antinociceptive action, and they have competitive effectiveness compared to classic analgesics. They may be chosen as the most effective from tested venoms for further development of pain relief remedies.

Cytotoxin antibody-based colourimetric sensor for field-level differential detection of elapid among big four snake venom

Development of a rapid, on-site detection tool for snakebite is highly sought after, owing to its clinically and forensically relevant medicolegal significance. Polyvalent antivenom therapy in the management of such envenomation cases is finite due to its poor venom neutralization capabilities as well as diagnostic ramifications manifested as untoward immunological reactions. For precise molecular diagnosis of elapid venoms of the big four snakes, we have developed a lateral flow kit using a monoclonal antibody (AB1; IgG1 – κ chain; Kd: 31 nM) generated against recombinant cytotoxin-7 (rCTX-7; 7.7 kDa) protein of the elapid venom. The monoclonal antibody specifically detected the venoms of Naja naja (p < 0.0001) and Bungarus caeruleus (p<0.0001), without showing any immunoreactivity against the viperidae snakes in big four venomous snakes. The kit developed attained the limit of quantitation of 170 pg/μL and 2.1 ng/μL in spiked buffer samples and 28.7 ng/μL and 110 ng/μL in spiked serum samples for detection of N. naja and B. caeruleus venoms, respectively. This kit holds enormous potential in identification of elapid venom of the big four snakes for effective prognosis of an envenomation; as per the existing medical guidelines.

Animal Studies on Venom Neutralization potential of Metal-Herbal (Copper-Leucas zeylanica) Nanocomposites (MHNC) Against Naja naja and Bungarus Caeruleus Venoms

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.

Elucidating the Venom Diversity in Sri Lankan Spectacled Cobra (Naja naja) through De Novo Venom Gland Transcriptomics, Venom Proteomics and Toxicity Neutralization

Inadequate effectiveness of Indian antivenoms in treating envenomation caused by the Spectacled Cobra/Indian Cobra (Naja naja) in Sri Lanka has been attributed to geographical variations in the venom composition. This study investigated the de novo venom-gland transcriptomics and venom proteomics of the Sri Lankan N. naja (NN-SL) to elucidate its toxin gene diversity and venom variability. The neutralization efficacy of a commonly used Indian antivenom product in Sri Lanka was examined against the lethality induced by NN-SL venom in mice. The transcriptomic study revealed high expression of 22 toxin genes families in NN-SL, constituting 46.55% of total transcript abundance. Three-finger toxins (3FTX) were the most diversely and abundantly expressed (87.54% of toxin gene expression), consistent with the dominance of 3FTX in the venom proteome (72.19% of total venom proteins). The 3FTX were predominantly S-type cytotoxins/cardiotoxins (CTX) and α-neurotoxins of long-chain or short-chain subtypes (α-NTX). CTX and α-NTX are implicated in local tissue necrosis and fatal neuromuscular paralysis, respectively, in envenomation caused by NN-SL. Intra-species variations in the toxin gene sequences and expression levels were apparent between NN-SL and other geographical specimens of N. naja, suggesting potential antigenic diversity that impacts antivenom effectiveness. This was demonstrated by limited potency (0.74 mg venom/ml antivenom) of the Indian polyvalent antivenom (VPAV) in neutralizing the NN-SL venom. A pan-regional antivenom with improved efficacy to treat N. naja envenomation is needed.

Autonomic Dysfunction Following Neurotoxic Snake Envenomation: Does Site of Bite Predispose?

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.