海水养殖真鲷最小弧菌外毒素的免疫学特性初步研究
THE IMMUNOLOGICAL CHARACTERISTICS OF THE EXOTOXIN Vm-Pm PRODUCEDBY VIBRIO MIMICUS
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摘要: 按常规方法免疫健康家兔进行外毒素抗体的制备。将制备的兔抗血清与等体积的外毒素混合,37℃下作用1h后,观察毒素的溶血性及细胞毒性,取免疫前家兔血清(零号血清)作阴性对照,用毒素作阳性对照。结果表明,兔抗毒素血清能中和毒素,抑制毒素对人血细胞的溶血性及对Vero细胞的细胞毒性,中和毒素的兔抗毒素血清最高稀释度为1∶256,毒素对照显示出溶血性及细胞毒性。毒素与一定浓度的嗜水气单胞菌HEC毒素抗血清、霍乱肠毒素抗血清在37℃下分别作用1 h后,观察其溶血性及细胞毒性。结果表明,这2种抗血清都不能中和毒素,毒素与这2种抗血清作用后,仍然保持其溶血性及细胞毒性。用浓度为0.2%的福尔马林对浓度为5mg/mL的外毒素进行灭活处理,得到的灭活疫苗为毒素苗。一定稀释度的毒素苗,在初次免疫真鲷2周后利用病原菌对免疫组及对照组的实验鱼进行人工攻毒感染,毒素苗的免疫保护率可达80%,强化免疫后,免疫保护率有所提高。注射免疫的免疫保护率比浸泡免疫的高。Abstract: The culture of red sea bream(Pagrus major) has undergone rapid development in recent years in China,however,suc- cessful production is increasingly hampered by diseases.Vibriosis,caused by Vibrio mimicus,has become a major cause of serious economical losses in the farming of red sea bream along the coast of southern China.Vm-Pm toxin was successfully purified from the culture supernatant of Vibrio mimicus strain 96-6-3,using sequential ammonium sulfate precipitation and column chromatogra-phy on DEAE-cellulose and Sephadex G100. In the present paper,the immunological characteristics of Vm-Pm toxin were stud-ied. Rabbits were immuned with three kinds of exotoxins to raise the anti-sera.After the exotoxins reacted with the anti-sera,their haemolytic and cytotoxic activities were examinated. The results showed that the Vm-Pm toxin was neutralized by anti-sera,then lost its haemolytic and cytotoxic activities.On the contrary,the HEC toxin and cholera enterotoxin still kept their haemolytic and cytotoxic activities after neutralization. Red sea breams (P. major)were injected intramuscularly and direct immersion withforma-linized Vm2Pm toxin(toxoid). The effectiveness of vaccination against V. mimicus was evaluated two weeks after the immuniza2 tion. The results showed that stronger protection was induced in red sea bream by toxoid.After booster immunization,this protec-tion was enhanced.
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Keywords:
- Vm-Pmtoxin /
- HEC toxin /
- Cholera enterotoxin /
- Toxid
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