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Subject: Animal Biochemistry × Author: DEKA, SURAKSHA SUBEDI × End date: 2017 × Start date: 2015 × Thesis Type: Ph.D ×
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    ThesisItemOpen Access
    EVALUATION OF EFFICACY OF WHOLE OUTER MEMBRANE PROTEIN OF Salmonella Typhimurium ADJUVANTED WITH CALCIUM PHOSPHATE NANOPARTICLES AS VACCINE CANDIDATE AGAINST SALMONELLOSIS IN CHICKEN
    (Assam Agricultural University, Khanapara, Guwahati, 2015-06) DEKA, SURAKSHA SUBEDI; Sarma, Satya
    Salmonella sp can infect a broad array of animals, causing diseases ranging from gastroenteritis to life threatening systemic infections. Substantial economic loss is manifested through mortality and poor growth of infected birds. Salmonella enterica serovar Typhimurium is the most frequently isolated serovar causing global food-borne outbreaks and chicken alone accounts for 10.4% of the total isolates. The control of salmonellosis can be accomplished either by vaccination or medication. Antibiotic resistance and issue of antibiotic residue is a major hurdle in medication. Thus control through immunization is the most efficient and economic method. The present study was conducted to study the efficacy of whole outer membrane protein adjuvanted with calcium phosphate nanoparticles (CAP-OMP) as vaccine candidate against salmonellosis in chicken. The study was carried out in Kamrupa birds which are dual purpose breed. The OMP was extracted from Salmonella Typhimurium (MTCC - 98) strain and confirmed by SDS-PAGE. CAP-OMP vaccine was then synthesized by method that is based on co-precipitation of calcium phosphate and OMP. The amount of protein entrapped in the complex was determined and the formulation was used to immunise the chicks on 14th day of life followed by booster dose after two weeks. The humoral immune response of the target vaccine was compared with aluminium hydroxide adjuvanted OMP and unadjuvanted OMP by indirect ELISA. Blood was collected from all the birds at 0 before vaccination and days 7, 14, 21, 28, 35 45 and 60 after booster vaccination. Some of the birds from each group were challenged on 60th day with either S. Typhimurium or with S. Gallinarum. The humoral response of the unchallenged birds was studied from 60th to 90th day. The CAP-OMP vaccine was able to elicit significantly higher antibody titres compared to other two groups up to 60th day post booster vaccination. However antibody titre level fell thereafter in all the groups. Mortality was not observed in any of the challenged birds but some clinical symptoms were exhibited by the control group. The challenged birds were slaughtered after one week and total CFU of either S. Typhimurium or S. Gallinarum per gram of liver was determined. It was observed that none of the vaccines gave total protection against challenge organisms but the Salmonella isolates recovered from the birds immunized with target vaccine was significantly lower than that of control group. The other two formulations viz; aluminium hydroxide –OMP vaccine and unadjuvanted OMP vaccine also decreased the bacterial count in liver tissue. Some important serum biochemical parameters were also studied from the vaccinated birds and it was found that the total protein and globulin content was significantly higher in CAP-OMP vaccinated group than the control groups. Creatine kinase level was significantly higher during first 21 days post booster vaccination birds given Aluminium hydroxide -OMP indicating some tissue damage at the site of injection caused by Aluminium hydroxide. Other parameters like alkaline phosphatase and creatinine showed non-significant fluctuations throughout the experiment. From this study, it can be concluded that CAP-OMP (S.Typhimurium) vaccine can be an effective vaccine candidate against salmonellosis.