Open Access
Research
(Published
online: 12-09-2016)
8.
First detection of canine
parvovirus type 2b from diarrheic dogs in Himachal Pradesh -
Shalini Sharma, Prasenjit Dhar, Aneesh Thakur, Vivek Sharma
and Mandeep Sharma
Veterinary World, 9(9): 964-969
doi:
10.14202/vetworld.2016.964-969
Shalini Sharma :
Department of
Veterinary Microbiology, Dr. G. C. Negi College of Veterinary
and Animal Sciences, Chaudhary Sarwan Kumar Himachal Pradesh
Krishi Vishvavidyalaya, Palampur - 176 062, Himachal Pradesh,
India; sharmashalini096@gmail.com
Prasenjit Dhar :
Department of
Veterinary Microbiology, Dr. G. C. Negi College of Veterinary
and Animal Sciences, Chaudhary Sarwan Kumar Himachal Pradesh
Krishi Vishvavidyalaya, Palampur - 176 062, Himachal Pradesh,
India; thakurprasen727779@gmail.com
Aneesh Thakur :
Department of
Veterinary Microbiology, Dr. G. C. Negi College of Veterinary
and Animal Sciences, Chaudhary Sarwan Kumar Himachal Pradesh
Krishi Vishvavidyalaya, Palampur - 176 062, Himachal Pradesh,
India; aneesh.thakur@gmail.com
Vivek Sharma :
Department of
Microbiology, College of Basic Sciences, Chaudhary Sarwan Kumar
Himachal Pradesh Krishi Vishvavidyalaya, Palampur - 176 062,
Himachal Pradesh, India; ankvivek@gmail.com
Mandeep Sharma :
Department of
Veterinary Microbiology, Dr. G. C. Negi College of Veterinary
and Animal Sciences, Chaudhary Sarwan Kumar Himachal Pradesh
Krishi Vishvavidyalaya, Palampur - 176 062, Himachal Pradesh,
India; mandeepsharma289@gmail.com
Received: 25-05-2016, Accepted: 05-08-2016, Published online:
12-09-2016
Corresponding author:
Prasenjit
Dhar, e-mail: thakurprasen727779@gmail.com
Citation:
Sharma S, Dhar P, Thakur A, Sharma V, Sharma M (2016) First
detection of canine parvovirus type 2b from diarrheic dogs in
Himachal Pradesh, Veterinary World, 9(9): 964-969.
Abstract
Aim:
The present
study was conducted to detect the presence of canine parvovirus
(CPV) among diarrheic dogs in Himachal Pradesh and to identify
the most prevalent antigenic variant of CPV based on molecular
typing and sequence analysis of VP2 gene.
Materials and
Methods:
A total of 102
fecal samples were collected from clinical cases of diarrhea or
hemorrhagic gastroenteritis from CPV vaccinated or
non-vaccinated dogs. Samples were tested using CPV-specific
polymerase chain reaction (PCR) targeting VP2 gene, multiplex
PCR for detection of CPV-2a and CPV-2b antigenic variants, and a
PCR for the detection of CPV-2c. CPV-2b isolate was cultured on
Madin-Darby canine kidney (MDCK) cell lines and sequenced using
VP2 structural protein gene. Multiple alignment and phylogenetic
analysis was done using ClustalW and MEGA6 and inferred using
the Neighbor-Joining method.
Results:
No
sample was found positive for the original CPV strain usually
present in the vaccine. However, about 50% (52 out of 102) of
the samples were found to be positive with CPV-2ab PCR assay
that detects newer variants of CPV circulating in the field. In
addition, multiplex PCR assay that identifies both CPV-2ab and
CPV-2b revealed that CPV-2b was the major antigenic variant
present in the affected dogs. A PCR positive isolate of CPV-2b
was adapted to grow in MDCK cells and produced characteristic
cytopathic effect after 5th
passage. Multiple sequence alignment of VP2
structural gene of CPV-2b isolate (Accession number HG004610)
used in the study was found to be similar to other sequenced
isolates in NCBI sequence database and showed 98-99% homology.
Conclusion:
This
study reports the first detection of CPV-2b in dogs with
hemorrhagic gastroenteritis in Himachal Pradesh and absence of
other antigenic types of CPV. Further, CPV-specific PCR assay
can be used for rapid confirmation of circulating virus strains
under field conditions.
Keywords:
canine parvovirus, molecular typing, phylogenetic analysis,
sequencing.
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