Isolation and adaptation of bovine herpes virus Type 1 in embryonated chicken eggs and in Madin-Darby bovine kidney cell line
| Autor: | Fateh V. Singh, Varsha Rani Gilhare, Nidhi Rawat, Sanjay Shakya, Devprabha Samrath |
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| Rok vydání: | 2015 |
| Předmět: |
0301 basic medicine
animal structures 040301 veterinary sciences Madin–Darby bovine kidney cell line animal diseases Veterinary medicine viruses Semen Biology SF1-1100 Virus cytopathic effect 0403 veterinary science 03 medical and health sciences SF600-1100 Cytopathic effect General Veterinary urogenital system Embryonated chorioallantoic membrane 04 agricultural and veterinary sciences biochemical phenomena metabolism and nutrition Ouchterlony double immunodiffusion Virology Animal culture Immunodiffusion 030104 developmental biology Cell culture pock biology.protein Bovine herpes virus Type 1 Antibody Research Article |
| Zdroj: | Veterinary World Veterinary World, Vol 9, Iss 2, Pp 222-225 (2016) |
| ISSN: | 0972-8988 |
| Popis: | Aim: Objective of the present study was to isolate bovine herpes virus Type 1 (BHV-1) from semen of infected bull and to adapt it onto embryonated eggs and Madin–Darby bovine kidney (MDBK) cell line. Further, the virus was identified by agar gel immunodiffusion (AGID) test. Materials and Methods: Semen samples were collected from five BHV-1 positive bulls previously confirmed for the presence of antibodies against BHV-1 using avidin-biotin enzyme linked immunosorbent assay test. The virus from semen samples was adapted in chorioallantoic membrane (CAM) of 11-day-old embryonated chickens eggs and in MDBK cell line. The presence of BHV-1 in infected CAM and cell culture fluid was confirmed by AGID test. Results: Virus infected CAM showed edema, congestion and thickening at first passage level. Small foci ranged from 1 to 2 mm in diameter, scattered all over the membrane were observed at first passage. More severe changes were observed in CAM after serial passaging. The large pock lesions, round in shape with opaque raised edge and depressed gray central area of necrosis ranged from 3 to 5 mm in diameter were developed at fourth passage. Blind passages in MDBK cell culture were made. The MDBK cell line at second passage level showed characteristic cytopathic effect viz. rounding of cells with shrinkage, followed by aggregation or clumping of cells which progressed rapidly and appeared as “bunch of grapes” at 72 h post inoculation. Few cells become elongated when compared with uninfected controls. A homogenate of CAM with distinct pock lesions and infected cell culture fluid developed precipitation line within 48 h against specific anti-BHV-1 immune serum by AGID test. Conclusion: BHV-1 was easily adapted in CAM of chicken embryos and in MDBK cell line. Virus infected CAM and cell culture fluid showed precipitin band by AGID test. |
| Databáze: | OpenAIRE |
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