Alborzi Alireza, Jolodar Abbas, Seyfi Abad Shapouri Masoudreza, Bagherian Pour Esmaeil
Department of Pathobiology, School of Veterinary Medicine, Shahid Chamran University of Ahvaz, Ahvaz, Iran;
Department of Basic Sciences, School of Veterinary Medicine, Shahid Chamran University of Ahvaz, Ahvaz, Iran.
Vet Res Forum. 2014 Fall;5(4):307-11.
Oestrus ovis is an economically important parasite of small ruminants and a zoonotic parasite with many reports of ophthalmomyiasis in human from Iran and other countries. The aim of the peresent study was the isolation and identification of excretory-secretory (ES) and somatic (S) antigens of O. ovis second and third stage larvae (L2, L3) collected from Arabi sheep breeds located in southwest of Iran. Positive sera were prepared by marking the sheep, taking blood sample and direct observation of the parasite in the head. Somatic antigens of the larvae (SL2, SL3) were prepared by sonication. Larval excretory-secretory antigens (ESL2, ESL3) were prepared by incubation the larvae in RPMI-1640 RPMI medium. Electrophoretic protein profiles of ESL2 two, ESL3 seven, SL2 eight, SL3 fifteen bands (from 79.0 to below 14.4 KDa) were shown. In immunoblotting with positive sera, four common bands in SL2 and SL3 at 58, 42.0, 29.0 and 28.0 kDa, one specific band in SL3 at 47.0 kDa and one band in ESL2, at 28.0 kDa, and three bands in ESL3 at 58.0, 42.0, 29.0 and 28.0 kDa were recognized. Among the profiles, the 28 kDa protein was the most common antigenic component. Nevertheless, the antigenic proteins 29, 58 kDa were a common protein in electrophoretic patterns of both S and ES proteins of L2 and L3 but, 42.0 kDa antigen the only one detected in immunoblot but not in S and ES protein profiles of the larvae. Therefore, the antigens 29.0, 42.0 and 58.0 kDa can be used for further studies of protective effects and serological diagnostic methods.
羊狂蝇是小型反刍动物的一种具有经济重要性的寄生虫，也是一种人畜共患寄生虫，在伊朗和其他国家有许多关于人类眼蝇蛆病的报道。本研究的目的是分离和鉴定从位于伊朗西南部的阿拉伯绵羊品种采集的羊狂蝇第二和第三期幼虫（L2、L3）的排泄分泌（ES）和虫体（S）抗原。通过标记绵羊、采集血样并直接观察头部的寄生虫来制备阳性血清。幼虫的虫体抗原（SL2、SL3）通过超声处理制备。幼虫排泄分泌抗原（ESL2、ESL3）通过将幼虫在RPMI - 1640培养基中孵育制备。显示了ESL2的两条、ESL3的七条、SL2的八条、SL3的十五条带（79.0至低于14.4 kDa）的电泳蛋白图谱。在用阳性血清进行免疫印迹时，在SL2和SL3中识别出58、42.0、29.0和28.0 kDa的四条共同条带，在SL3中识别出47.0 kDa的一条特异性条带，在ESL2中识别出28.0 kDa的一条条带，在ESL3中识别出58.0、42.0、29.0和28.0 kDa的三条条带。在这些图谱中，28 kDa蛋白是最常见的抗原成分。然而，29、58 kDa的抗原蛋白是L2和L3的S和ES蛋白电泳图谱中的共同蛋白，但42.0 kDa抗原是在免疫印迹中唯一检测到但不在幼虫的S和ES蛋白图谱中的蛋白。因此，29.0、42.0和58.0 kDa的抗原可用于进一步研究保护作用和血清学诊断方法。
