de la Fe Christian, Assuncão Patricia, Ramírez Ana S, Poveda Jose B
Unit of Epidemiology and Preventive Medicine, Veterinary Faculty, University of Las Palmas, Spain.
Berl Munch Tierarztl Wochenschr. 2004 Jan-Feb;117(1-2):1-5.
The suitability of 5 agents for the inactivation of different field strains of the four mycoplasma species associated with contagious agalactia syndrome in goats, i.e. Mycoplasma agalactiae, Mycoplasma mycoides subsp. mycoides LC, Mycoplasma capricolum subsp. capricolum and Mycoplasma putrefaciens, was investigated. Immunoprophylaxis of this syndrome is still based on inactivated vaccines, which traditionally use formalin as the inactivating agent. Moreover, the limited information existing about this type of vaccine is only based on assays against Mycoplasma agalactiae. Our results showed that formalin (0.1%, 37 degrees C during 16 hours) and phenol (0.5%, 24 hours) were effective against all species tested. Surprisingly, binary ethileneimine (BEI), a classical virus-inactivating agent, also proved to be very effective when it was used in a 0.1 M concentration over 24 hours. With heat treatment, every species was inactivated at 60 degrees C. No satisfying results were obtained with purified saponin. To evaluate the harmful effects of each agent on mycoplasmal proteins, a representative strain was subjected to an effective inactivation protocol with each agent, which was monitored by Western immunoblotting. Immunoblotting was performed using sera of animals inoculated with the respective mycoplasma species, to compare the effect of all the agents on treated strains with untreated strains. The results confirmed that phenol, BEI and to a lesser extent also formalin inactivated all species without causing a significant damage while heat caused stronger damage on surface proteins. Future in vivo studies should be conducted because, as recently shown, the combined use of a suitable inactivant and adjuvant could give rise to the induction of certain cytokines and strong antibody production of a specific isotype pattern, thus opening ways to develop more efficacious inactivated vaccines against contagious agalactia.
研究了5种试剂对与山羊传染性无乳综合征相关的4种支原体不同田间菌株的灭活适用性，这4种支原体分别为无乳支原体、丝状支原体丝状亚种、山羊支原体山羊亚种和腐败支原体。该综合征的免疫预防仍基于灭活疫苗，传统上使用福尔马林作为灭活剂。此外，关于这类疫苗的现有信息有限，且仅基于针对无乳支原体的检测。我们的结果表明，福尔马林（0.1%，37℃处理16小时）和苯酚（0.5%，处理24小时）对所有测试的支原体均有效。令人惊讶的是，经典的病毒灭活剂双乙烯亚胺（BEI）在0.1 M浓度下处理24小时时也被证明非常有效。经热处理，每种支原体在60℃时均被灭活。纯化皂苷未取得满意结果。为了评估每种试剂对支原体蛋白质的有害影响，用每种试剂对一个代表性菌株进行有效的灭活方案处理，并通过蛋白质免疫印迹法进行监测。使用接种了相应支原体物种的动物血清进行免疫印迹，以比较所有试剂对处理菌株和未处理菌株的影响。结果证实，苯酚、BEI以及程度较轻的福尔马林可灭活所有支原体物种，且不会造成显著损伤，而热处理对表面蛋白造成的损伤更大。由于最近的研究表明，合适的灭活剂和佐剂联合使用可诱导某些细胞因子，并产生特定同种型模式的强抗体反应，从而为开发更有效的抗传染性无乳综合征灭活疫苗开辟道路，因此未来应进行体内研究。
