Proteomic and functional characterisation of Trimeresurus popeiorum (Pope's pit viper) venom proteins: Role of enzymatic and non-enzymatic venom toxins in envenomation pathophysiology.

Snakebite remains a significant public health issue in tropical regions, with 4.5 to 5.4 million incidents annually. Trimeresurus popeiorum (Pope's Pit Viper), found in Southeast Asia and northeast India, poses a potential threat, yet its venom's protein composition and toxicity are poorly understood. In this study, we used label-free quantitative proteomics to analyze the venom of T. popeiorum, identifying 106 proteins across 12 venom protein families. Notably, 60 % of the venom consisted of proteolytic enzymes, correlating with its prominent metalloprotease, fibrin(ogen)lytic, procoagulant, and thrombin-like activities. The proteome composition also correlates with the clinical effects such as consumption coagulopathy and local effects, seen in victims of Pit Viper envenomation in northeast India. Our findings suggest that T. popeiorum venom is less toxic than other Viperinae species such as Daboia russelii and Echis carinatus, likely due to isoform-level variations in certain toxin classes, including metalloprotease and serine protease. The venom's lethal dose (LD50) in Swiss albino mice was 1 mg/kg, and it caused haemorrhage, tissue necrosis, edema, myotoxicity, and defibrinogenation. Histopathological examination of the TPV-treated mice showed notable toxic effects, including marked hepatic vacuolation in the liver, damage to cardiac muscle and vascular congestion in the heart, bronchial epithelial hyperplasia with cellular infiltration in the interstitial and peribronchiolar regions of the lungs, as well as tubular necrosis and haemorrhage in the kidneys. This research provides the first comprehensive analysis of T. popeiorum venom, highlighting its pharmacological effects and the need for greater medical attention to this lesser-known species.