Thesis (Ph.D.) - Cairo University - Faculty of Veterinary Medicine - Department of Microbiology

Mycobacterium avium subsp. paratuberculosis (MAP) is the cause of Johne's disease (JD), a chronic granulomatous intestinal disease of cattle. The disease represents one of the important problemsfacing dairy industry andcausing high economic losses. 100 fecal and 100serum samples were collected from JD clinically suspected cases.Sixtyout of the 100 fecal samples werepositive usingZiehl Neelsen stain showing acid and alcohol fast red bacilli.. Bacteriological examination of the 100 fecal samples reveled the isolation of 34MAP isolates.The recovered MAPisolates were tested by conventional and nested PCR.The nested PCR provedthe identity of the 34 MAP isolates; however, the conventional PCR using different primers gave variable results. Using a commercialMAP specific ELISAkits, only 13 out of the 100 serum samples were positive. The recoveredMAP isolates were cultured on modified Middlebrook7H9 and modified Watson Reid liquid media for preparation of theMAP cultural filtrate (CF) proteins, which were then separated by SDS-PAGE.The MAPCF immunogenic determinants were identified using western-Blot technique. Different immunogenic bands from 9-72 kDa were identified

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