A functional and thromboelastometric-based micromethod for assessing crotoxin anticoagulant activity and antiserum relative potency against Crotalus durissus terrificus venom
Research article published in Toxicon (2018)
Abstract
The assessment of the capacity of antivenoms to neutralize the lethal activity of snake venoms still relies on traditional rodent in vivo lethality assay. ED50 and LD50 assays require large quantities of venoms and antivenoms, and besides leading to animal suffering. Therefore, in vitro tests should be introduced for assessing antivenom neutralizing capacity in intermediary steps of antivenom production. This task is facilitated when one key lethal toxin is identified. A good example is crotoxin, a β-neurotoxin phospholipase A2-like toxin that presents anticoagulant activity in vitro and is responsible for the lethality of venoms of Crotalus durissus snakes. By using rotational thromboelastometry, we reported recently one sensitive coagulation assay for assessing relative potency of the anti-bothropic serum in neutralizing procoagulant activity of Bothrops jararaca venom upon recalcified factor-XII-deficient chicken plasma samples (CPS). In this study, we stablished conditions for determining relative potency of four batches of the anti-crotalic serum (ACS) (antagonist) in inactivating crotoxin anticoagulant activity in CPS (target) simultaneously treated with one classical activator of coagulation (agonists). The correlation coefficient (r) between values related the ACS potency in inactivating both in vitro crotoxin anticoagulant activity and the in vivo lethality of whole venom (ED50) was 0.94 (p value < 0.05). In conclusion, slowness in spontaneous thrombin/fibrin generation even after recalcification elicit time lapse sufficient for elaboration of one dose-response curve to pro- or anti-coagulant agonists in CPS. We propose this methodology as an alternative and sensitive assay for assessing antivenom neutralizing ability in plasma of immunized horses as well as for in-process quality control.
Abstract sourced from PubMed (NCBI) for the cited record. See the original publication for the authoritative version.
Resumen
A functional and thromboelastometric-based micromethod for assessing crotoxin anticoagulant activity and antiserum relative potency against Crotalus durissus terrificus venom.
Por qué esto importa para la hirudoterapia
Este estudio estableció un micrométodo tromboelastométrico rotacional utilizando plasma de pollo deficiente en factor XII recalcificado para evaluar la actividad anticoagulante de la crotoxina y determinar la potencia neutralizante relativa de cuatro lotes de suero anticrotálico frente al veneno de Crotalus durissus. Los valores in vitro de neutralización anticoagulante correlacionaron con los datos in vivo de letalidad ED50 (r = 0,94; p < 0,05), lo que respalda el método como ensayo alternativo para evaluar la capacidad neutralizante del antiveneno en plasma de caballos inmunizados y para el control de calidad en proceso. Para el ámbito de ASH, la relevancia es muy indirecta: el trabajo concierne a la actividad anticoagulante de la crotoxina, una toxina similar a fosfolipasa A2 del veneno de serpiente, y a la evaluación de antivenenos, sin involucrar sanguijuelas, sustancias derivadas de sanguijuelas ni hirudoterapia. El único vínculo conceptual es el dominio compartido de la biología de la coagulación sanguínea, y los hallazgos no tienen aplicabilidad directa en la investigación o la práctica de la terapia con sanguijuelas.
Citación
A functional and thromboelastometric-based micromethod for assessing crotoxin anticoagulant activity and antiserum relative potency against Crotalus durissus terrificus venom
Prezoto BC et al. · Toxicon, 2018
Contexto clínico relacionado
Explore cómo esta investigación se conecta con la práctica clínica
Añadido a la biblioteca ASH: May 27, 2026 · Última actualización del sitio: 18 de junio de 2026