THE IMMUNOCHEMICAL REACTIVITY AND NEUTRALIZING CAPACITY OF POLYVALENT Vipera (EUROPEAN) ANTIVENOM ON ENZYMATIC AND TOXIC ACTIVITIES IN THE VENOMS OF CROTALIDS FROM ARGENTINA

Abstract

The immunochemical reactivity and neutralizing capacity of polyvalent Vipera antivenom (Vipera ammodytes, Vipera aspis, Vipera berus, Vipera lebetina, and Vipera xanthina) were tested on the enzymatic and biological activities of Crotalus durissus terrificus and the following Bothrops venoms from Argentina (Bothrops alternatus, Bothrops ammodytoides, Bothrops neuwiedii, Bothrops jararaca, Bothrops jararacussu, and Bothrops moojeni). The Vipera antivenom reacted weakly when tested by double immunoprecipitation (DIP) and reacted with all the venoms when tested by ELISA. Several components in all the venoms studied were recognized in Western blots. Vipera antivenom deactivated to different degrees in vitro procoagulant, (indirect) hemolytic, and proteolytic activities in all the venoms studied. Preincubation of Bothrops alternatus venom with Vipera antivenom neutralized a lethal potency of 4.5 LD50 in mice with an ED50 of 1.25 <FONT FACE="Symbol">±</FONT> 0.25 <FONT FACE="Symbol">m</FONT>l per <FONT FACE="Symbol">m</FONT>g of venom, and with 1.0 <FONT FACE="Symbol">m</FONT>l/<FONT FACE="Symbol">m</FONT>g inhibited 54% of the hemorragic activity and 48% of necrotic activity. Vipera antivenom (2.0 <FONT FACE="Symbol">m</FONT>l per <FONT FACE="Symbol">m</FONT>g toxin) inhibited the phospholipase A2 activity of purified crotoxin and decreased its lethal potency by 60%, while the neutralizing capacity on the lethal potency of crude Crotalus durissus terrificus venom was poor even at a level of 5.0 <FONT FACE="Symbol">m</FONT>l/<FONT FACE="Symbol">m</FONT>g of venom

Application of the 3R Concept in the Production of European Antiviperinum on Horses – Multisite, Low Volumes Immunization Protocol and Elisa

“…Improving antiviperinum production by immunizing horses with horned viper toxin is still important because horned viper is the most venomous and most widespread European venomous snake. In addition, V. ammodytes antivenom is cross-reactive with the venom of the common European adder (V. berus), as well as with venoms of other European Viperidae [26,27]. In this paper, we describe a new immunization protocol used for anti-viper antivenom production.…”

Comparative assay of Vipera ammodytes antivenom potency

“…One of the most used and aged antivenom potency assessing method is based on the injection in the animal experiment bodies of a significant venom-antivenom mixture and interpolation of the 50% survival animal 1,3,7,8,10,11 . Another way is represented by immunoenzymatic method for the assay of antivenom potency [11][12][13][14] . In these cases, antivenom potency is directly articulated on venom unit quantity but, as it is already well known, even the venom of the same snake species differs (sometime substantially) in theirs biological and toxicological properties.…”

“…Because all venoms (particularly harvested from snake) possess a very significant enzymatic content, the antivenom potency can be also determined by antienzymatic assay 1,13,15,16 . The antivenom efficacy values obtained by all previously mentioned methods can be indirectly compared using correlation coefficients 15,16 .…”