Abstract
There are currently many controversial issues in the study of bovine respiratory syncytial infection. In this regard, it is relevant to study the biological properties of the virus, optimize the methods of its cultivation and select the most technologically advanced methods of designing diagnostic and prevention tools for this disease. The aim of this work was to select sensitive cell systems and to optimize the cultivation parameters in selected cell cultures. The Vologda/2019 strain of the bovine respiratory syncytial infection virus isolated from biological material obtained from a calf with respiratory symptoms was used in the experiment. The strain was adapted to the continuous cell culture derived from bovine turbinate tissue (BT) and deposited in the State collection of microorganism strains at FGBI “ARRIAH”. It was established that the continuous cell lines of fetal bovine trachea (FBT) and calf kidney (RBT) are the most sensitive cell systems for the reproduction of the bovine respiratory syncytial virus strain Vologda/2019, the maximum accumulation of the virus was observed in these cell cultures. The cytopathic activity of the virus in the FBT cell culture ranged from 4.78 ± 0.18 to 5.50 ± 0.16 lg TCID 50 /cm 3 , and in the RBT cell culture – from 4.00 ± 0.23 to 4.75 ± 0.20 lg TCID 50 /cm 3 at days 4–5 of cultivation. It was determined that in case of multiplicity of inoculation of FBT and RBT cell cultures with the virus at 0.1 lg TCD 50 /cell and the use of 2% glutamine in the maintenance nutrient medium, as well as 2% horse or cattle blood serum, it is possible to obtain virus material with high cytopathic activity.
Highlights
It is relevant to study the biological properties of the virus, optimize the methods of its cultivation and select the most technologically advanced methods of designing diagnostic and prevention tools for this disease
The Vologda/2019 strain of the bovine respiratory syncytial infection virus isolated from biological material obtained from a calf with respiratory symptoms was used in the experiment
The strain was adapted to the continuous cell culture derived from bovine turbinate tissue (BT) and deposited in the State collection of microorganism strains at FGBI“ARRIAH”
Summary
В работе использовали вирус РСИ КРС штамма «Вологда/2019», выделенный из биологического материала, полученного от теленка с признаками респираторной патологии, и адаптированный к перевиваемой линии культуры клеток слизистой носовых перегородок КРС (ВТ) с титром цитопатической активности 4,0 lg ТЦД50/см. В культуру клеток инокулировали вирус РСИ КРС штамма «Вологда/2019» в дозе 0,1 ТЦД50/кл. Инокуляцию вируса проводили после удаления ростовой питательной среды, далее на 1,5 ч оставляли вирус для контакта с клеточным монослоем в CO2инкубаторе при температуре 37 °С, а затем добавляли поддерживающую полусинтетическую питательную среду (ПСС) с содержанием в ней по 2% сыворотки крови животных (коровы или лошади) и глютамина. После контакта вируса и клеточного монослоя добавляли 0,1 см питательной среды ПСС с содержанием в ней по 2% сыворотки крови лошади и глютамина. Учет результатов титрования вируса проводили через 10 сут инкубирования при условии сохранения целостности монослоя клеток в контрольных лунках [12]. 4. Культура клеток RBT без инокуляции вируса РСИ КРС (5-е сут, увеличение ×200). РСИ КРС в ИФА (ELISA kit for antigenic diagnosis of Bovine Respiratory Syncytial Virus (BRSV); Bio-X Diagnostics, Бельгия) согласно инструкции производителя
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