Experimental model for removal of snake venom via hemoperfusion in rats

Abstract

ObjectiveTo examine the efficiency of hemoperfusion in removing South American rattlesnake (Crotalus durissus terrificus) venom from rats compared with neutralization by antivenom.DesignAn exploratory experimental investigation in rats involving the injection of snake venom with or without subsequent hemoperfusion or antivenom administration.SettingBasic animal research laboratory in a private university.AnimalsNormal, healthy male Wistar rats (0.29‐0.40 kg, 3‐6 months old) from a commercial breeder.InterventionsFour experimental groups of randomly allocated rats (n = 3/group) were studied: Group 1: rats were injected with a single dose of venom (5 mg/kg, IM, in the right thigh) with no other treatment; blood samples were collected minutes before death to determine leukocyte, platelet, and erythrocyte counts; Group 2 (Control): rats underwent hemoperfusion alone for 60 min using a hemoperfusion cartridge designed for protein adsorption (by granulated charcoal) and protein precipitation (by tannic acid); Group 3 (Venom + antivenom): rats were injected with venom (5 mg/kg, IM) and, 10 min later, were treated with antivenom at the venom:antivenom ratio recommended by the manufacturer; Group 4 (Venom + hemoperfusion): Rats were injected with venom (5 mg/kg, IM) and, 10 min later, were hemoperfused for 60 min. In groups 2‐4, blood samples were collected for leukocyte, platelet, and erythrocyte counts 24 h after venom.Measurements and Main ResultsRats injected with venom alone (Group 1) developed signs of neurotoxicity and ataxia and died in 9.0 ± 0.43 h but showed no changes in leukocyte or erythrocyte counts. In contrast, there were no deaths in groups 2‐4. The lack of deaths in Groups 3 and 4 indicated that antivenom and hemoperfusion, respectively, protected against the lethal effects of the venom.ConclusionsHemoperfusion with a double‐action hemoperfusion cartridge capable of protein adsorption and precipitation protected rats against C. d. terrificus venom.

Evaluation of Protection by Caffeic Acid, Chlorogenic Acid, Quercetin and Tannic Acid against the In Vitro Neurotoxicity and In Vivo Lethality of Crotalus durissus terrificus (South American Rattlesnake) Venom

“…The samples were prepared in 2× standard buffer (65.8 mM Tris-HCl, 26.3% glycerol, 2.1% SDS and 0.01% bromophenol blue) containing 0.5% β-mercaptoethanol (Bio-Rad) at 100 • C for 10 min. The gel was run in a vertical Mini-Protean electrophoresis system (150 V, 30 mA, 15 W, 45 min), stained for 30 min with Coomassie Brilliant Blue G-250 (Bio-Rad) and washed in deionized water, essentially as described elsewhere [8,64]. The amount of venom to be used was calculated based on a rat weighing 400 g. Thus, 2.4 mg of venom or 50 mg of tannic acid were each dissolved in 1 mL of phosphate-buffered saline (PBS).…”

“…Envenomation by the South American rattlesnake (Crotalus durissus terrificus) is characterized by minimal or no local manifestations, such as pain, mild edema, and recurrent erythema, but marked systemic effects, including coagulopathy, myotoxicity with myoglobinuria, peripheral neuromuscular blockade (flaccid paralysis), and acute kidney injury (AKI) [1][2][3][4][5][6][7][8]. AKI is the principal cause of death [1,4,6], with the interval between envenomation and antivenom administration being an important factor in determining AKI and mortality [6,9].…”