Project description:This project mainly aims to characterize the complex toxic components present in the venom of Trimeresurus malabaricus (Malabar pit viper). Since Trimeresurus malabaricus (Malabar pit viper) species are mainly inhabited to plantation crop areas, its envenomation is a serious threat to the human population thriving in these zones, especially to the plantation workers. Therefore, exploring the venom proteome of Malabar pit viper is decisive to develop and design new antivenom and therapeutics against its envenomation. As described in this study, applying various orthogonal separation strategies helped in dissecting venom constituents of Trimeresurus malabaricus and is the first comprehensive attempt in revealing the complex venom profile of Malabar pit viper through proteomics approaches incorporating multiple database searches. In order to achieve this the crude venom components were resolved on a 12% SDS page. Further each of the bands were subjected to in-gel trypsin digestion. The crude venom was also subjected to ion-exchange chromatography separation. The obtained fractions were subjected to in-solution trypsin digestion. All the digested peptides were then subjected to Q-TOF LC-MS/MS analysis.
Project description:Trimeresurus gracilis is a pit viper inhabiting the high mountains of Taiwan. No specific antivenom against T. gracilis is available, and we did not find any prior published case reports on its clinical envenomation. We present two patients with T. gracilis envenomation who were both bitten on the index finger and were administered bivalent hemotoxic antivenom against Trimeresurus stejnegeri and Protobothrops mucrosquamatus. In case 1, the patient was administered seven vials of antivenom within 2 days of envenomation. She received surgical intervention on day 26 and recovered from the wound 3 months after envenomation, but her left index finger was slightly and irreversibly bent. In case 2, the patient was administered 12 vials of antivenom within 10 hours after envenomation, received surgical intervention on day 1, and underwent debridement and reconstruction surgery on day 7 after envenomation. The wounds healed 1.5 months after envenomation, and no deformity occurred. Laboratory data showed elevated D-dimer levels and prothrombin times. The cross-neutralizing ability of bivalent hemotoxic antivenom against T. stejnegeri and P. mucrosquamatus appeared insufficient to treat the local effects of T. gracilis envenomation. The deformity of the finger of the patient in case 1 might have been caused by the lower dose, later administration of antivenins, and a delay in the necessary surgery.
| S-EPMC8991321 | biostudies-literature
Project description:short read WGS of Trimeresurus albolabris
Project description:BackgroundThe venom of bamboo vipers (Trimeresurus stejnegeri - TS), commonly found in Taiwan, contains deadly hemotoxins that cause severe envenomation. Equine-derived antivenom is a specific treatment against snakebites, but its production costs are high and there are some inevitable side effects. The aim of the present work is to help in the development of an affordable and more endurable therapeutic strategy for snakebites.MethodsT. stejnegeri venom proteins were inactivated by glutaraldehyde in order to immunize hens for polyclonal immunoglobulin (IgY) antibodies production. After IgY binding assays, two antibody libraries were constructed expressing single-chain variable fragment (scFv) antibodies joined by the short or long linker for use in phage display antibody technology. Four rounds of biopanning were carried out. The selected scFv antibodies were then further tested for their binding activities and neutralization assays to TS proteins.ResultsPurified IgY from egg yolk showed the specific binding ability to TS proteins. The dimensions of these two libraries contain 2.4 × 107 and 6.8 × 107 antibody clones, respectively. An increase in the titers of eluted phage indicated anti-TS clones remarkably enriched after 2nd panning. The analysis based on the nucleotide sequences of selected scFv clones indicated that seven groups of short linkers and four groups of long linkers were identified. The recombinant scFvs showed significant reactivity to TS venom proteins and a cross-reaction to Trimeresurus mucrosquamatus venom proteins. In in vivo studies, the data demonstrated that anti-TS IgY provided 100% protective effects while combined scFvs augmented partial survival time of mice injected with a lethal amount of TS proteins.ConclusionChickens were excellent hosts for the production of neutralization antibodies at low cost. Phage display technology is available for generation of monoclonal antibodies against snake venom proteins. These antibodies could be applied in the development of diagnostic kits or as an alternative for snakebite envenomation treatment in the near future.
Project description:The two venomous pit vipers, Trimeresurus macrops and T. hageni, are distributed throughout Thailand, although their abundance varies among different areas. No species-specific antivenom is available for their bite victims, and the only recorded treatment method is a horse antivenom raised against T. albolabris crude venom. To facilitate assessment of the cross-reactivity of heterologous antivenoms, protein profiles of T. macrops and T. hageni venoms were explored using mass-spectrometry-based proteomics. The results show that 185 and 216 proteins were identified from T. macrops and T. hageni venoms, respectively. Two major protein components in T. macrops and T. hageni venoms were snake venom serine protease and metalloproteinase. The toxicity of the venoms on human monocytes and skin fibroblasts was analyzed, and both showed a greater cytotoxic effect on fibroblasts than monocytic cells, with toxicity occurring in a dose-dependent rather than a time-dependent manner. Exploring the protein composition of snake venom leads to a better understanding of the envenoming of prey. Moreover, knowledge of pit viper venomics facilitates the selection of the optimum heterologous antivenoms for treating bite victims.