Tasima Lídia J, Serino-Silva Caroline, Hatakeyama Daniela M, Nishiduka Erika S, Tashima Alexandre K, Sant'Anna Sávio S, Grego Kathleen F, de Morais-Zani Karen, Tanaka-Azevedo Anita M
Laboratory of Herpetology, Butantan Institute, São Paulo, SP, Brazil.
Interinstitutional Postgraduate Program in Biotechnology (PPIB - IPT, IBU and USP), University of São Paulo(USP), São Paulo, SP, Brazil.
J Venom Anim Toxins Incl Trop Dis. 2020 Apr 6;26:e20190053. doi: 10.1590/1678-9199-JVATITD-2019-0053.
is considered one of the most important species of venomous snakes in Brazil, due to the high mortality of its snakebites. The venom of contains four main toxins: crotoxin, convulxin, gyroxin and crotamine. Venoms can vary in their crotamine content, being crotamine-negative or -positive. This heterogeneity is of great importance for producing antivenom, due to their different mechanisms of action. The possibility that antivenom produced by Butantan Institute might have a different immunorecognition capacity between crotamine-negative and crotamine-positive venoms instigated us to investigate the differences between these two venom groups.
The presence of crotamine was analyzed by SDS-PAGE, western blotting and ELISA, whereas comparison between the two types of venoms was carried out through HPLC, mass spectrometry analysis as well as assessment of antivenom lethality and efficacy.
The results showed a variation in the presence of crotamine among the subspecies and the geographic origin of snakes from nature, but not in captive snakes. Regarding differences between crotamine-positive and -negative venoms, some exclusive proteins are found in each pool and the crotamine-negative pool presented more phospholipase A than crotamine-positive pool. This variation could affect the time to death, but the lethal and effective dose were not affected.
These differences between venom pools indicate the importance of using both, crotamine-positive and crotamine-negative venoms, to produce the antivenom.
由于其蛇咬伤导致的高死亡率，被认为是巴西最重要的毒蛇种类之一。的毒液包含四种主要毒素：响尾蛇毒素、convulxin、gyroxin和响尾蛇胺。毒液中的响尾蛇胺含量可能不同，分为响尾蛇胺阴性或阳性。由于它们不同的作用机制，这种异质性对于生产抗蛇毒血清非常重要。Butantan研究所生产的抗蛇毒血清在响尾蛇胺阴性和阳性毒液之间可能具有不同免疫识别能力的可能性促使我们研究这两种毒液组之间的差异。
通过SDS-PAGE、蛋白质印迹法和ELISA分析响尾蛇胺的存在，而通过HPLC、质谱分析以及抗蛇毒血清致死率和效力评估对两种毒液进行比较。
结果表明，自然环境中的蛇的亚种和地理来源之间响尾蛇胺的存在存在差异，但圈养蛇中没有差异。关于响尾蛇胺阳性和阴性毒液之间的差异，在每个样本池中发现了一些独特的蛋白质，并且响尾蛇胺阴性样本池中的磷脂酶A比响尾蛇胺阳性样本池更多。这种差异可能会影响死亡时间，但致死剂量和有效剂量不受影响。
毒液样本池之间的这些差异表明，使用响尾蛇胺阳性和阴性毒液来生产抗蛇毒血清都很重要。
