Investigation of bovine coronavirus strain CV-315 cultural properties

A. Berezenko, F. Vabishchevych, O. Godovskyі, V. Nedosekov
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Abstract

The purpose of this study was to investigate the features of cultivation of bovine coronavirus strain CV-315 isolated in Ukraine from a calf with coronavirus infection and to select optimal methods of virus cultivation to obtain viral material with the highest possible titers of infectious activity in order to develop manufacturing technology of means of immunoprophylaxis and specific diagnostics. During the study, the influence of a number of factors on the accumulation of strain CV-315 was studied: the presence and concentration of trypsin in the nutrient medium, the effect of fetal bovine serum, the degree of cell culture monolayer during virus infection, also the virus dose, temperature and the term of cultivation. According to the results, it was established that bovine coronavirus strain CV-315 has the highest infectious activity when cultured for 72 hours before the manifestation of CPE of 70–80%, without the addition of trypsin and fetal bovine serum content of 2% at 37±0.5°C. It was also found that the optimal infective dose is 0.1–0.01 viral particles per cell for infection of the fully formed monolayer of MDBK cell culture. The obtained results will be used in the development of veterinary vaccines against bovine coronavirus.
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牛冠状病毒CV-315株培养特性的研究
本研究旨在探讨乌克兰分离的牛冠状病毒CV-315株的培养特点,并选择最佳的病毒培养方法,以获得具有最高感染活性的病毒材料,从而开发免疫预防和特异性诊断手段的制造技术。在研究过程中,研究了多种因素对CV-315菌株积累的影响:营养培养基中胰蛋白酶的存在和浓度、胎牛血清的影响、病毒感染过程中细胞培养单层的程度、病毒剂量、温度和培养时间。结果表明,牛冠状病毒CV-315株在37±0.5℃条件下,在不添加胰蛋白酶和胎牛血清含量为2%的条件下,培养72 h后感染活性最高,CPE表现为70-80%。MDBK细胞培养的完全形成的单层细胞感染的最佳感染剂量为每细胞0.1 ~ 0.01个病毒颗粒。获得的结果将用于开发抗牛冠状病毒的兽医疫苗。
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