The Center for the Study of Venoms and Venomous Animals, UNESP, Botucatu, Sao Paulo, Brazil.
J Toxicol Environ Health A. 2010;73(13-14):926-33. doi: 10.1080/15287391003745069.
Equine antivenom is considered the only treatment for animal-generated envenomations, but it is costly. The study aimed to produce Apis mellifera (Africanized honeybee) and Crotalus durissus terrificus (C.d.t.) antivenoms using nanostructured silica (SBA-15) as adjuvant and cobalt-60 ((60)Co)-detoxified venoms utilizing young sheep. Natural and (60)Co-irradiated venoms were employed in four different hyperimmunization protocols. Thus, 8 groups of 60- to 90-d-old sheep were hyperimmunized, enzyme-linked immunosorbent assay (ELISA) serum titers collected every 14 d were assessed clinically daily, and individual weight were measured, until d 84. Incomplete Freund's (IFA) and nanostructured silica (SBA15) adjuvants were compared. The lethal dose (LD(50)) for both venoms was determined following intraperitoneal (ip) administration to mice. High-performance liquid chromatography on reversed phase (HPLC-RP) was used also to measure the (60)Co irradiation effects on Apis venom. At the end of the study, sheep were killed in a slaughterhouse. Kidneys were histologically analyzed. LD(50) was 5.97 mg/kg Apis and 0.07 mg/kg C.d.t. for native compared to 13.44 mg/kg Apis and 0.35 mg/kg C.d.t. for irradiated venoms. HPLC revealed significant differences in chromatographic profiles between native and irradiated Apis venoms. Native venom plus IFA compared with SBA-15 showed significantly higher antibody titers for both venoms. Apis-irradiated venom plus IFA or SBA-15 displayed similar antibody titers but were significantly lower when compared with native venom plus IFA. Weight gain did not differ significantly among all groups. (60)Co irradiation decreased toxicity and maintained venom immunogenic capacity, while IFA produced higher antibody titers. SBA-15 was able to act as an adjuvant without producing adverse effects. Hyperimmunization did not affect sheep weight gain, which would considerably reduce the cost of antiserum production, as these sheep were still approved for human consumption even after being subjected to hyperimmunization.
马用抗蛇毒血清被认为是治疗动物源性蛇咬伤的唯一方法,但价格昂贵。本研究旨在使用纳米结构化二氧化硅(SBA-15)作为佐剂,并用钴-60((60)Co)对蜂毒(非洲化蜜蜂)和响尾蛇毒(C.d.t.)进行解毒,并利用小羊进行制备。天然和(60)Co 照射的毒液被用于四种不同的超免疫方案。因此,8 组 60-90 日龄的绵羊被超免疫,每隔 14 天收集酶联免疫吸附测定(ELISA)血清滴度,每天进行临床评估,并测量个体体重,直到第 84 天。比较了不完全弗氏佐剂(IFA)和纳米结构化二氧化硅(SBA15)佐剂。通过腹腔(ip)给药,确定两种毒液的半数致死剂量(LD(50))。还使用反相高效液相色谱(HPLC-RP)测量(60)Co 照射对蜂毒的影响。研究结束时,绵羊在屠宰场被杀死。对肾脏进行组织学分析。与天然毒液相比,LD(50)为 5.97mg/kg 的 Apis 和 0.07mg/kg 的 C.d.t.,而照射后的 Apis 和 C.d.t.毒液分别为 13.44mg/kg 和 0.35mg/kg。HPLC 显示天然和照射后的 Apis 毒液在色谱图上存在显著差异。天然毒液加 IFA 与 SBA-15 相比,两种毒液的抗体滴度均显著升高。Apis 照射毒液加 IFA 或 SBA-15 显示出相似的抗体滴度,但与天然毒液加 IFA 相比,抗体滴度显著降低。所有组之间的体重增加没有显著差异。(60)Co 照射降低了毒液的毒性并保持了其免疫原性,而 IFA 产生了更高的抗体滴度。SBA-15 可以作为佐剂而不产生不良反应。超免疫不会影响绵羊的体重增加,这将大大降低抗血清的生产成本,因为这些绵羊在经过超免疫后仍可用于人类消费。