Open Access
Research
(Published
online: 27-02-2016)
20.
Isolation and adaptation of bovine herpes
virus Type 1 in embryonated chicken eggs and in Madin–Darby
bovine kidney cell line -
Devprabha Samrath, Sanjay Shakya, Nidhi Rawat, Varsha Rani
Gilhare and Fateh Singh
Veterinary World, 9(2): 222-225
doi:
10.14202/vetworld.2016.222-225
Devprabha Samrath:
Department of Veterinary Microbiology, College of Veterinary
Science and Animal Husbandry, Anjora Durg, Chhattisgarh, India;
devprabhavet@gmail.com
Sanjay Shakya:
Department of Veterinary Public Health, College of Veterinary
Science and Animal Husbandry, Anjora Durg, Chhattisgarh, India;
shakyadurg@gmail.com
Nidhi Rawat:
Department of Veterinary Microbiology, College of Veterinary
Science and Animal Husbandry, Anjora Durg, Chhattisgarh, India;
dr_nidhirawat@yahoo.com
Varsha Rani Gilhare:
Department of Veterinary Microbiology, College of Veterinary
Science and Animal Husbandry, Anjora Durg, Chhattisgarh, India;
docvarsha20@gmail.com
Fateh Singh:
Division of Animal Health, ICAR-Central Sheep and Wool Research
Institute, Avikanagar-304501, Rajasthan, India; fateh.ars07@gmail.com
Received: 03-06-2015, Accepted: 05-01-2016, Accepted:
16-01-2016, Published online: 27-02-2016
Corresponding author:
Nidhi Rawat, e-mail: dr_nidhirawat@yahoo.com
Citation:
Samrath D, Shakya S, Rawat N, Gilhare VR, Singh F (2016)
Isolation and adaptation of bovine herpes virus Type 1 in
embryonated chicken eggs and in Madin–Darby bovine kidney cell
line,
Veterinary World, 9(2):
222-225.
Abstract
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.
Keywords:
Bovine herpes virus Type 1, chorioallantoic membrane, cytopathic
effect, Madin–Darby bovine kidney cell line, pock.
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