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A definitive diagnosis of M. bovis infection in animals depends on the isolation of the organism, as the optimum condition for the primary isolation of M. bovis infection has not been determined. In this study a total number of 2550 cattle from different governorates in Egypt were tested by Single Intradermal Cervical (SID) tuberculin skin test. Positive reacted cattle (no= 42, 1.6%) were slaughtered and suspected lesions were collected, these samples were decontaminated by using three different methods, which were 0.75 % Hexadecyl Pyridinium Chloride (HPC), 5% oxalic acid and 6 % sulphuric acid. Isolation of mycobacteria from 42 samples by using these decontaminated methods were of 34 (80.9 %), 30 (71.4 %) and 28 (66.6 %), respectively. So it could be concluded that using HPC method as decontaminated agent revealed high rate of isolation of mycobacteria with less contamination (7.1 %) than the other two methods.

A B S T R A C TThe objective of the present work is to prepare and evaluate the effectiveness of a locally prepared polyvalent vaccine against mastitis from the most common causes of mastitis. Staphylococcus aureus, Streptococcus agalactiae and Escherichia coli were the most prevalent bacteria recovered from clinical and subclinical mastitis. Montanide ISA-206 adjuvanted inactivated polyvalent vaccine containing the three strains was prepared. Twenty pregnant cows were inoculated intramuscularly with the prepared polyvalent vaccine two months prior to calving and boostering at day 21 from the primary injection. Serum samples from vaccinated and non-vaccinated cows were collected at the 1 st , 3 rd , 8 th , 12 th , 16 th , 20 th and 24 th weeks post vaccination and evaluated immunologically using ELISA. The results showed that immune response was significantly higher in the vaccinated group than that of controls. These results could be indicated the safety and effectiveness of the vaccine in reduction of incidence and severity of clinical cases of mastitis but further studies should be done to elucidate the possibility of field application and effectively.