Development of a protocol for isolation and identification of snake toxins in plasma samples from patients victims of snakebite in Brazil

• Grant number: 24/14131-4
• Support Opportunities: Scholarships abroad - Research Internship - Doctorate
• Start date: March 01, 2025
• End date: August 31, 2025
• Field of knowledge: Biological Sciences - Biochemistry - Molecular Biology
• Principal Investigator: Rui Seabra Ferreira Junior
• Grantee: Joeliton dos Santos Cavalcante
• Supervisor: Alejandro Alagon
• Host Institution: Centro de Estudos de Venenos e Animais Peçonhentos (CEVAP). Centro Virtual de Toxinologia. Universidade Estadual Paulista (UNESP). Campus de Botucatu. Botucatu , SP, Brazil
• Institution abroad: Universidad Nacional Autónoma de México (UNAM), Mexico
• Associated to the scholarship: 22/16060-1 - From mechanisms to biomarkers of envenoming by Brazilian snakes: a multicentric study for the development of strategies in diagnosis and monitoring, BP.DR

Abstract

The complexity of snakebite envenoming makes diagnosis a major challenge in clinical practice. By employing state-of-the-art proteomics on blood plasma from snake-envenomated patients with different clinical conditions, we are exploring the mechanisms involved in the pathophysiology of snakebite envenoming. However, the difficulty in differentiating accidents caused by different species when the snake is not identified, as in cases of snakebite by Bothrops moojeni and Crotalus durissus terrificus, is an obstacle. The presence of common manifestations, such as edema and coagulation changes, in both cases, makes it difficult to identify the type of accident in regions where these species overlap, which can lead to false negative diagnoses and follow-ups. This uncertainty leads to delays in antivenom treatment, during which time the disease can progress from mild to moderate or severe. The need to monitor creatine kinase levels in the first 12 hours to differentiate snakebite and the late onset of neurological manifestations in Crotalus cases are examples of how differential diagnosis can be challenging. This delay can result in more serious clinical complications, the need for a greater number of antivenom vials, and an increased risk of adverse reactions. The present proposal aims to develop a protocol for the isolation and identification of circulating toxins in samples from patients victims of snakebite in Brazil. In addition to quantifying the main families of toxins present in the samples at the time of hospital admission. We will use plasma samples from patients victims of snakebite by B. atrox, B. moojeni and C. d. terrificus and we will employ a multidisciplinary approach using ELISA assays, coupling of antibodies to dynobeads and isolation of toxins from samples, mass spectrometry and bioinformatics. This collaboration between the Center for the Study of Poisons and Venomous Animals at Universidade Estadual Paulista Júlio de Mesquita Filho (CEVAP/UNESP) and the National Autonomous University of Mexico will allow the development of an innovative protocol for identifying toxins in clinical samples, deepening understanding of venomic variability and its clinical impacts. The results generated from this plan of activities abroad and carried out within the scope of the Regular FAPESP Proc. 2022/16060-1, in addition to generating candidate biomarkers for the diagnosis of snakebite, will allow the production of more accurate diagnostic kits and the understanding of the mechanisms that lead to different clinical manifestations. Finally, the exchange of knowledge during the period will allow for initial training, testing of research instruments and analysis models, paving the way for a proposal for international cooperation with the aim of continuing studies.