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Abstract In this study a total of 360 (nasal,buccal and ocular swabs; lung, tracheal and lymph node)samples collected from apparently healthy , diseased livingand slaughtered cattlewere used forisolation of Mycoplasma spp. Biochemical and serological identification of 67 isolates of Mycoplasma revealed presence of 5 different Mycoplasma spp. , the most prevalent one was M.bovis 40.2% followed by M.arginine 31.3%,M.bovirhinis 16.9%,M.boviculi 5.9% and finally M.bovigentalium 1.49%.Three M.bovisisolates were subjected to 6 differentantimicrobial(Enrofloxacin,Danofloxacin,Erythromycin,Florofenicol,Oxytetracycline,Tulathromycin) at different concentrations to determine the MIC using micro dilution method.MIC of the isolated M.bovis was ranged from 0.125-8μg/ml for quinolones, 8to≥128μg/ml for Erythromycin, ≥0.5 to ≥128μg/ml for macrolides , 4-64μg/ml for florofenicol and 2-64μg/ml for oxytetracycline.PCR technique was applied using specific primer targeting 16srRNA for Mycoplasma of ruminant, species identification through specific primers for M.bovis, M.arginine, M.bovirhinis.sequencing of the amplified product of M.bovis, M.arginine, M.bovirhinisand analysis of these sequences showed high similarity and identity with strains published on gene bank . Flouroquinolones resistance has been estimated through specific primers targets Gyrase A and par C in QRDR of the isolated M.bovisstrain. Amplified product of par C was sequenced and analysis of this sequence revealed highsimilarity with identity 100% and presence of mutations at different sites that changed produced amino acids that reflected on phenotype showing increased resistance. |