DETECTION OF ENTERIC DNA VIRUSES FROM DIARRHEIC DOGS OF HIMACHAL PRADESH
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Date
2019-07-20
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CSKHPKV. Palampur
Abstract
Diarrhea is among the most common clinical conditions seen in dogs leading to high morbidity
and mortality. It can have various causes, including infectious agents, like viruses, bacteria and
parasites. The aim of the study was to isolate enteric DNA viruses from diarrheic dogs of
Himachal Pradesh and characterize them at molecular level. A total of 238 fecal/other samples
were collected from diarrheic / apparently healthy dogs along with brief clinical history of the
animal. The faecal samples were subjected to DNA extraction and PCR for the detection of
enteric DNA viruses viz. CPV, CAV-1 and Dog CV with primers targeting the VP2, pⅦ and
replicase gene of CPV, CAV-1 and Dog CV respectively of each virus were used in the study.
109 (45.79 %) samples were found positive for CPV-2b and 4 (1.68%) samples were found
positive for CAV-1. No samples were found positive for Dog CV. HA along with HI and DotELISA were standardized for virus detection from clinical samples and infected cell culture fluids
using hyper immune sera raised in rabbits against CPV and CAV-1. Dot-ELISA was found to be
almost similar to in sensitivity HA-HI for detection of viruses. Growth studies of CPV and CAV-
1 were done on MDCK, A-72 and Vero cells. CPV could be adapted to MDCK and A-72 cells
but not to Vero cells while adaptation of CAV-1 was not possible on any cell culture.
Physicochemical characterization of CPV was done by checking the effect of temperature, pH,
chloroform and formalin on the virus. The VP2 (CPV-2) and pⅦ (CAV-1) gene fragment was
sequenced directly which revealed close genetic similarity to Indian and Italian isolates of CPV
and CAV, respectively. Microbiological studies on fecal samples revealed 37 bacterial isolates
that were resistanct to penicillin and streptomycin in antibiogram and were found concurrently
associated with clinical cases of viral diarrhea. In conclusion, diarrhea in dogs in H.P are mostly
caused by CPV type 2b strain and occasionally by CAV-1 and is frequently associated with other
bacterial species.