Master Thesis      [Total: 29 ]

Trials for the production of monoclonal antibodies against a local isolate of haemophilus paragallinarum

Roukaya Osman, 2004


one monoclonal antibody (MAb) was produced against a locally isolated H.paragallinarum serovar A strain which reacted serologically in indirect ELISA test and also reacted in Western blot test with a single band of 39 KDa which is the same band of haemagglutinin (HA) antigen. this MAb was used to differentiate between the standard strain by different tests, and it was found that it was reacted specifically with the W standard strain of serovar A and not reacted with strains of serovar B and C. From this study it is recommended to use this MAb for characterization of H.paragallinarum serotypes and selection of suitable strains for vaccine production instead of the traditional methods.

Factors affecting tuberculin production

Shereen Mohamed fouad, 2002


studies on the effect of some factors on the yield of tuberculin. First studies on the effect of types of media on the yield of PPD, showed that the growth of mycrobacteria in Dorset and henely's synthetic media was better than in Sauton or long's medi. Our studies showed that human isolated much rapid and better growth than bovine strain. Studies on the relationship between PH of culture filtrate and yield between 7-12 weeks, was associated with drop in Ph. Four different types of tuberculin either (human, bovine) were prepared by different methods which were prepared by different methods which weredissolved into two types of buffers one contain glycerol and other tween 80 -First type was prepared by inactivated the mycobacterium strains by heating and precipitation of protein by trichloroacetic acid solution 40%, this gave the best yield than other types -Second type was prepared by inactivated the isolated by phenol solution 1% and precipitated the protein as first type -Third type was prepared by inactivation with heating and precipitation of tuberculoprotein with ammonium sulfate solution 50% -Fourth type was prepared by inactivation the isolates with phenol 1 % and precipitationof tubercuoprotein with ammonium sulfate so this type of PPD gave lower yield, but the best one which gave clear diameter of tuberculin reaction. Finally the specificity differences were calculated as SPD = (Aa + Bb)-(Ab+ Ba)

Evaluation of some mycobacterial antigens for diagnosis of animal tuberculosis

Mohamed Osman, 2002


The present work comprises the isolation and purification of the 38KDa glycoprotein antigen from unheated culture filtrate of M.tuberculosis H37Rv strain. Also, this study aimed to comparison between the 38 KDa antigen and bovine PPD tuberculin for the diagnosis of tuberculosis in artificially infected guinea pigs and naturally infected buffaloes. First , M. tuberculosis H37Rv strain was cultivated on Middle Brook 7H9 medium for 6 weeks after which the glycoprotein content in this medium was precipitated by ammonium sulphate at concentration of 45 %. Second, the 38 KDa glycoprotein antigen was then purified by alcohol fractionation and the result was three precipitates and one supernatant. The obtained product was identified as a single band migrate at 38 KDa on SDS-PAGE. Reactivity of the antigen was tested by Western blot that gave non-photographable bands then tried Dot-blot which gave positive results with P2, P3, last supernatant differ according to concentration of antigen. Evaluation of the 38 KDa antigen versus bovine PPD tuberculin was done on infected and non infected guinea pigs with different strains of typical mycobacteria (M. tuberculosis, M.bovis) and a typical mycobacteria (M.avium, M.Kansasii, M. fortuitum and M. intracellulare) using skin tuberculin test and ELISA. The obtained results of tuberculin test revealed that the 38 KDa antigen over the bovine PPD tuberculin could distinguish between M. kansaii infected guinea pigs from those infected with M.tuberculosis complex. Additionally, ELISA results confirmed the superiority of the 38 KDa antigen over the bovine PPD tuberculin in distinguishing between guinea pigs infected with typical and a typical mycobacteria. Validity of using the 38 KDa as a specific antigen for diagnosis of bovine tuberculosis by ELISA was confirmed by testing of 15 tuberculin positive and five tuberculin negative buffaloes. ELISA positive results obtained with 8 buffaloes previously identified as M.bovis infected buffaloes and one was negative culture while negative results were recorded with both the 5 tuberculin negative and the other 6 tuberculin positive buffaloes in which 2 were previously identified as MOTT infected buffaloes, one gave negative culture, and three identified as M.bovis infected buffaloes a result which could not be fulfilled by bovine PPD tuberculin. The specify of the 38 KDa antigen was 88.9 % while that of bovine PPD was 66.7%. It was obvious from the outcome the superiority of 38 KDa over bovine PPD tuberculine in diagnosis of bovine tuberculosis in buffaloes.

Studies on some factors improving the production of antitetanic serum with high potency

Mahmoud Mohammed, 2001


Production of high potency antitetanic serum by addition of some adjuvant such as oil , aluminium hydroxide gel and some drugs such a levamisolle, ranitidine. the antibody titers wer evaluated by using serum neutralization test (snt). it was found that the inoculated animals with tetanus toxoid adjuvanted with oil exhibited a high level of antibodies then aluminium hydroxide gel and later levamisole.

Studies on some factors improving the production of antitenaic serum with high potency

Mahmoud Mohammed, 2001


Production of high potency antitetanic serum by addition of some adjuvant such as oil, aluminium hydroxide gel, and some drugs such as levamisole, Ranitidine. The antibody titres were evaluated by using serum neutralization (SNT). It was found that the inoculated animals with tetanus toxoid adjuvanted with oil exhibited a higher level of antibodies then aluminium hydroxide gel and later levamisole.

Studies on haemorrhagic viral disease in rabbits

Owis Salman, 1999


RVHD is still a great problem threaten rabbit industry in Egypt. RVHD could be diagnosed generally owing to host specificity. Very high mortality in adults, very short course of the disease, typical I.n. lesions especially necrotic pale liver and haemorrhagic tracheo pneumonic and specific histopathological finding. Haemagglutination test using human RBCs type “O” is still a satisfactory test for preliminary diagnosis of RVHD. The liver is the most suitable organ for RHDV detection. RHDV could be isolated in susceptible rabbits and failed to be isolated in SPF ECE and cell culture. RVHD is a highly contagious disease transmitted experimentally via contaminated feed and water, infected air and contact infection. The inactivated RHDV vaccine is simple to be manufactured and easy to be used and should be applied on rabbits beginning from 1.5 months old. The RHDV vaccine could be applied safely in pregnant ewes without abortions or still birth and proved very high success as an emergency vaccine during RVHD outbreak.

Immunological studies on the efficacy of some sheep pox strains in goat

Abd Elrazek Omar, 1999


Immunological studies on pasteurella multocida in cattle

Eman Ahmed Zaki, 1999


Eleven pasteurella multocida of cattle origin were confirmed, a correlation between biochemical and serotyping revealed four groups of strains and enzyme decapsulation confirmed the identity of these strains as belonging to type (B:2; B:2.5; B:3.4; B:1, -:7 AND -2.7). Electrophoretic protein profile revealed two groups of strains. The first group were similar to the standard (B:2) strain. The second group of strains were smilar to each other, DNA fingerprinting profiles indicate three groups. The strains varied in their virulence to mice and immunological comparison indicated homologous and some heterologus cross protection.

Study on trial for praparation of fowl pox vaccine locally on embryonated specific pathogen free eggs

Namaa Abd El-hafiz, 1998


Fowl pox (WP1) strain was paasaged to 6 passage on SPF embryonated egg and chicken embryo fibroblast (CEF)for vaccine preparation.The maximum titre of inoculated culture was around the 2oth hours and on the 4th day after ECE inoculation. The prepared vaccines were safe, sterile and the best temparture fro virus preservation was -20oC. Challenge of vaccinated birds revealed protection96% for egg propagated vaccine and 92% for CEF adapted vaccine. Seroconversion was done using ELISA and SNT to compare the efficiency of the locally prepared vaccines with imported commercial vaccines which revealed that the local egg propagated vaccine gave better immunogenicity than the imported and the tissue culture adapted one, so it is suitable for birds vaccination

A prespective study of borrelia anserina isolation and cultivation approach

lilian Saad, 1998


The procedure descibd in this work proved that Borrelia anserina ( spirochchaetes) could be cultivated success fully in vitro in two stabilized media ( stabilized borrelia growth medium "A' and stabilized BSKI . the chance of contaminating the medium is greatly reduced by addition of effective is possible to carry these spirochaetes in two successive subcultures.the infectivity of intial culture to chickens was 100%.the infectivity of culture is not lost by cultivation although, there is atendancy to become attenuated after the growth continued.