PREVALENCE AND MOLECULAR CHARACTERIZATION OF CANINE PARVOVIRUS FROM DIARRHOEIC DOGS

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dc.contributor.advisorJHALA, M. K.
dc.contributor.authorDHIRAJLAL, UMRETIYA SANDIP
dc.date.accessioned2018-05-25T07:26:06Z
dc.date.available2018-05-25T07:26:06Z
dc.date.issued2009
dc.description.abstractCanine parvovirus (CPV), a member of the genus Parvovirus, family Parvoviridae is a non-enveloped, single-stranded DNA virus of approximately 5 kb genome. The virus was first described in 1978, with the original isolate being termed CPV type 2 (CPV-2). In 1979, a variant CPV strain designated CPV type 2a (CPV-2a) became widespread, followed by further antigenic variants designated CPV type 2b (CPV-2b) and 2c (CPV-2c), which emerged in 1984 and 2000, respectively. In due course, CPV-2a, CPV-2b and 2c replaced CPV-2. The vaccines presently in use are CPV-2 strain specific. Hence, the objective of the present study was to detect and genotype CPV at different locations of Gujarat state by Polymerase chain reaction (PCR), using primer sequences derived from the conserved VP2 region of the genome, and to subsequent Restriction fragment length polymorphism (RFLP) analysis of the PCR product. The RFLP analysis employed the REs Hphl and Mboll in order to differentiate the CPV-2 antigenic variants. A few epidemiological factors like breed, age, sex, season and vaccination status affecting the infection were also studied. Haemagglutination (HA) and Haemagglutination inhibition (HI) tests were also employed so as to compare their sensitivity with PCR. From three different districts of Gujarat state, a total of 55 faecal samples were collected from dogs showing signs of diarrhea by sterile rectal swabs during December 2008 to April 2009. The samples were clarified and processed by HA-HI and PCR (using M1/M2 primers) assays for CPV detection. PCR detected the virus in 20 (36.36%) samples. Significantly more cases (40.91%) were seen in dogs below six months of age than dogs above six months of age (28.57%). Local breed showed maximum incidence (54.54%) followed by Doberman (42.85%), Labrador (41.66%), Alsatian (37.5%), Great Dane (33.33%) and Pomeranian (18.18%). Maximum number of cases (45.00%) was recorded in winter season i.e. during the months of December to February than in summer months (13.33%). Male dogs revealed higher incidence (41.05%) than female dogs (25.00%). CPV-unvaccinated dogs showed higher incidence (42.86%) of CPV than the CPV-vaccinated dogs (29.63%). HA and HI tests using pig erythrocytes yielded 17 (30.91%) samples positive for CPV with 85.00% sensitivity, 100%) specificity and 94.54 % overall agreement relative to PCR results. PCR amplification with primer pair M1/M2 yielded product of -252 bp in all 20 positive samples similar to that of vaccine (Puppy DP, Intervet) strain and the specificity of PCR amplicons was confirmed by RFLP using RE Fast Digest Dral. Nine representative PCR positive samples (representing six breeds of dog) were processed further for genotyping study. Primer pair Pabs/Pabas amplified all selected nine positive faecal samples with the product of 682 bp, but the vaccine strain could not be amplified suggesting that the strain present in faecal samples was not CPV-2 and was a variant. The specificity of PCR amplicons was confirmed by RFLP using RE Fast Digest Hinji. PCR amplification with primer pair Pbs/Pbas yielded the product (428 bp) specific to primers used suggesting that the strain in faecal samples was not CPV-2a but some other variant strain. Primer pair Hfor/Hrev used earlier in one study could not amplify any of the nine CPV positive faecal samples, but the vaccine strain was amplified, while primer pair Hfor/Pbas could amplify all nine CPV positive samples and vaccine strain yielding expected product (915 bp) specific to the primer pair used. Digestion of PCR product amplified by primer pair Hfor/Pbas with RE Hphl differentiated field strain of CPV from vaccine strain i.e. CPV-2. PCR products amplified by primer pairs Pbs/Pbas and Hfor/Pbas were digested by RE MboII, which detects Glu-426 mutation at nt 4062-4064 and hence, all nine CPV positive faecal samples and recognized as CPV-2c.en_US
dc.identifier.urihttp://krishikosh.egranth.ac.in/handle/1/5810047547
dc.keywordsPREVALENCE AND MOLECULAR CHARACTERIZATION, CANINE PARVOVIRUS, DIARRHOEIC DOGSen_US
dc.language.isoenen_US
dc.publisherAAU, Ananden_US
dc.research.problemPREVALENCE AND MOLECULAR CHARACTERIZATION OF CANINE PARVOVIRUS FROM DIARRHOEIC DOGSen_US
dc.subVeterinary Microbiologyen_US
dc.subjectVERTINARY MICROBIOLOGYen_US
dc.subjectCHARACTERIZATIONen_US
dc.themePREVALENCE AND MOLECULAR CHARACTERIZATION OF CANINE PARVOVIRUS FROM DIARRHOEIC DOGSen_US
dc.these.typeM.V.Sc.en_US
dc.titlePREVALENCE AND MOLECULAR CHARACTERIZATION OF CANINE PARVOVIRUS FROM DIARRHOEIC DOGSen_US
dc.typeThesisen_US
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