الفهرس 
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Abstract
Current study aim is the : Prepation of combined MG and MS in one inactivated vaccine with nanoparticles (IMS1313) adjuvants. MontanideTM IMS 1313 N VPR (IMS 1313) adjuvant consists of awater-dispersed liquid nanoparticles as emulsion combined with an immunostimulating compound. Because this adjuvant has an aqueous phase, it is suitable as a mucosal delivery vehicle .
This performed by :
1- Mycoplasma gallisepticum and Mycoplasma synoviae were identified on culture media and biochemically then genotypic identification by PCR and sequencing on gene bank
2- These isolates propagated in SPF to increase its virulence , Inactivated by lysis buffer and binary ethylene amine
3- The prepared vaccine subjected to different quality control tests including sterility, safety and potency tests. Results indicate that the success of the vaccine to be satisfactory to all quality control parameters.
4- Evaluation of prepared vaccine through challenge test were delivered via intranasal and S/C routes of administration in 775 of 14 day old Cobb breed chicks chickens which divided into 8 groups:
5- Cell mediated and humoral immune responses were measured.
The result revealed that :
The result of Challenge test against M. gallisepticum and M.synoviae vaccination revealed that, chicken groups vaccinated with single dose of locally prepared combined lysis MG and MS vaccine and challenged with virulent M. gallisepticum R-strain and virulent MS strain showed the highest degree of protection. Which against MS give 96 % I/N and 92% S/C and against MG give 88 % I/N and 84 % S/C while chicken groups vaccinated with single dose of locally prepared combined binnary MG and MS vaccine given s/c challenged with virulent M. gallisepticum R-strain virulent MS strain showed good degree of protection ,which against MS give 92% I/N and 88 % S/C and against MG give 84% I/N and 80 % S/C while commercial killed MG vaccine gave 76%protection against MG, Then chicken groups vaccinated with single dose of commercial live MG vaccine and challenged with virulent M. gallisepticum R-strain gave 84%protection against MG. while chicken groups vaccinated with single dose of commercial live MS vaccine and challenged with virulent MS give 80 %protection against MS.
2 -Evaluation of cytokine ( IL4,IL6 and IFNγ) after vaccination: Through all Over mean reveald that group of chicken vaccinated by locally prepared combined lysis MS and MG vaccine induced high response at 3,5,7,10, 15 and 21 days post vaccination then group of chicken vaccinated live commercial MS vaccine then group of chicken vaccinated locally prepared combined binnary MS and MG vaccine then group of chicken vaccinated live commercial MG vaccine and reached the highest response at 21 days post vaccination
3- Assessment lymphocyte proliferation assay were showed gradual enhancement and the highest activity was 21th day for all groups and group vaccinated by locally prepared combined lysis vaccine was the higher group then groupvaccinated by locally prepared combined binnary vaccine then group vaccinated by live commercial MS. ,Finally group vaccinated by live commercial MG, consequently.
4- Monitoring of the humoral immune response developed against M. gallisepticum and M.synoviae, it can clearly be noticed that a rapid rise of antibody titer starting from 1st week . The peak of antibody titer achieved at 6weeks post vaccination. Both locally prepared combined MG and MS vaccine ethier by lysis or binary used in vaccination of chickens under this study were more efficient in the protection of chickens against M. synoviae and M.gallisepticum respectively which gave the highest titre of antibody than commercial MG vaccine ,but locally prepared combined lysis MG and MS vaccine is higer in antibody titre than locally prepared combined binary MG and MS vaccine against MS&MG. it can clearly be noticed that a rapid rise of antibody titer was observed after vaccination with a booster dose.
All the results were compared with that of SPF chicks non-vaccinated kept as negative control that had no macrophage activity.
In conclusion: Combined vaccines have the advantage of providing protection against more than one disease causing the same symptoms, reducing vaccination cost and number of infections per farm as well as saving time and reducing the stress reactions. Inactivation of Mycoplasma by Lysis and production of lysate bacteria (particulate antigen) give the higer immune response and protection .The results of the present study document the immuno enhancing effects of Montanide adjuvants on immune response of vaccinated birds and demonstrate the potentiality nanoparticles adjuvantes when used via inactivated vaccine through intra nasal route. Mass application of such vaccines will be add value to improve the vaccination strategies against Avian Mycoplasmosis.