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Kerala Veterinary and Animal Sciences University, Wayanad

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1993 - 19992
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Subject: Veterinary Microbiology × End date: 1999 × Start date: 1993 × Thesis Type: Ph.D ×
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    ThesisItemOpen Access
    PRODUCTION AND APPLICATION OF MONOCLONAL ANTIBODIES AGAINST DUCK PLAGUE VIRUS
    (College of Veterinary and animal Science,Mannuthy, 1999) RAVINDRA DATTATRAYA PADALKAR; V. Jayaprakasan
    MonocloiMl iinlibodies (Mabs) were raised against the vaccine strain of DPV and three strains of DPV Mz, Vaccine (OPV^V). IVRl (DPV-1) and Alleppy strain (DPV-A) were used to raise polyclonal serum in tite present investigation. DPV-V was revived in 11 day old chicken enibiyo and the enibrjo deatli was recorded four to five days PI with congestion all over the body and spleen and nccrotic foci in liver. The cytopathy in CEF cell culture observed was rounding and clumping of the cells, syncytium formation and bridge fonriation with extensive vacuolation m the Cytoplasm. The detachment of the cells was observed at 120 h PI. 1)PV-1 a virulent strain was inocnlaied in the ducklings, death was recorded in all the inoculated birds with extensive hemorrhages on serous membranes, muscles and visceral organs. Nccrotic foci on li\ei, enlargement and congestion of liver and spleen, and white necrotic foci m the gizzard were evident. The virus was further passaged in DDE and cultivated in bulk in DEP cell culture. The Dl'V-V and Dl'V-A were titrated in Clil' cell cultnie and the TCin,o was 4.7 X 10' per nd of the inoeulnm lor DPV-V and 3.2 X lO' for I)PV-A. ni'V-i cultivated in Did' cell culture had a IC 11),,i of the inoculum All the strains were partially purified at 100000 g for 4.5 h at 4" C in Heekman ultra centrifuge and the protein concentration of the virus was estimated by biuret method and was found to he 1 1 mg. 8 mg and 7 mg for DPV-V, A and I respectively. All the three strains of DPV were inoculated in mice to raise polyclonal serum. Four mice out of five inoculated with DPV-V showed FJdSA titres more than 1:12800, one mouse showed a litre of 1:6400. The mice inoculated witi* DPV-A showed a litre of more than 1:12800 and those inoculated with DPV-I, 1:6400 FddSA was used to test the sera samples of the miee inoculated with DPV strains. The test was found to be highly sensitive, easy to perlorm and less time consuming. The test therefore can be recommended for routine diagnosis of DPV I'olyclonal seiuin was used to study ll'.c cross reactivitv of the three strains oC I)PV with FIJSA. DI'V-V polyclonal scrum reacted with the liomologous virus with a titre of 1:12800. It also showed similar titer with other two heterologous strains. I'olyclonal serum raised against DI'V-A had a titre of 1:12800 with homologous and hcterologuus strains of DI'V. DPV-I reacted with Immnhigous strain at a titre uf 1:6400 Similar titrcs were observed with hertologous strains. Immiino pero.xidase test was used for the detection of the tissue antigens in DPV infected CEF monolayers and in liver and spleen sections. Polyclonal serum raised against DPV-V detected homologous virus in the CF.F monolayers and hetrologous virus in the liver and spleen sections. The staining reaction was observed as dark brown deposits at the virus localization. However background tissue was also stained brown to faint yellow in the stained preparation. The virus neutralization test was used to study the cross neutralization by employing polyclonal .serum raised against the three strains of DPV. Polyclonal serurTi raised against DPV-V showed a VN F of 64 with a VNl of 1.8 with homologous virus and a VN F 45 with VNI 1.65 with other two strains of DPV. Polyclonal serum raised against DPV-A neutralized the homologous virus at a titre of 32 with a VNl of 1.5. it also neutnrlized the vacciiic strain nf DI'V with same VNT and VNI, However it neutralized DPV-1 with VN I' 24 and VNI 1.35. Dl'V-i poiyclonal serum neutralized the homologous virus at a titre of 45 with a VNI of 1.65 . I he s.iiue neutralized DPV-V and DPV-A with a VN T 32 and VNI 1.5.
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    ThesisItemOpen Access
    ASSESSMENT OF IMMUNITY TO DUCK PLAGUE VIRUS (DUCK VIRUS ENTERlTlS) ON VACCINATION
    (COLLEGE OF VETERINARY AND ANIMAL SCIENCES-MANNUTHY,THRISSUR, 1993) DIWAKAR DATTATRAYRAO KULKARNI; P.c. James
    During 1991, six outbreaks clinically duck plague (DP) with 33 per cent morbidity mortality were investigated. Duck plague from each outbreak. The isolates were able lesions and death of the duck embryos but fai chicken embryos during initial passages. suspected to be and 26 per cent was isolated to produce the led to kill the One of the strain, named DP-S was par by 10 passages in chicken embryos following duck embryos. Though the attenuated strain did pathogenicity index was reduced from 1.9 to 1. DP-S under transmission electron microscope of herpes virus morphology. Two DP vaccines - commercial vaccine vaccine having virus titres 0.74 and 3.5 respectively, were separately inoculated into ducklings respectively, two groups receiving two receiving double dose of corresponding interval of four weeks. Another group of duckl as control without vaccination. tially attenuated 20 passages in kill ducks, its 23. The isolate revealed virions and lab-adapted log 10 ELD 50/ml four groups of single dose and vaccines at an ings was kept (ii) challenged with Thre e ducks in each group were virulent DPV at four,eight and 20 weeks post-vaccination. The birds in all the five groups were screened at regular intervals for studying the immune response by virus neutralization (VN), leucocyte migration-inhibition (LMI) and passive haemagglutination (PHA) test. The challenged and survived birds were the carrier status of DPV by examination of their for virus isolation. In an organized farm, 180 ducks were gi vaccine at one year of age and were scre antibodies, LMI response and PHA titres before post-vaccination. Randomly selected two birds six weeks post-vaccination. The findings of the study are briefly l Six duck plague outbreaks were investigat isolated, and characterized. It was in duck and chicken embryos. partially The commercial vaccine could elicit very response as compared to laboratory adapted immunity could not last long even upto ei single vaccination and 20 weeks in double screened for rectal swabs ven commercial ened for VN and eight weeks were challenged isted as under: ed, the virus attenuated poor immune vaccine. The ght weeks in vaccination. (iii) * Single vaccination is not effective as c vaccination given four weeks apart. The assessment of antibody-mediated (AMI) and cell mediate d (CMI) immune responses indicated that both and CMI are involved in protection of ducks against duck plague. The vaccinated or vaccinated and infected show carrier status as attempts to isolate rectal swabs collected after vaccination were unsuccessful. The PHA has been standardized for diagnosis of duck plague.