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Abstract A total of 617 sample from 38 poultry farm and small backyards located in different sources in El-Kalyobia , El-Monofia and El-sharkia Governorate, during the period of January 2016 to September 2019. All samples were phenotypically screened for presence of different causative microorganisms. • Bacteriological examination revealed 102 Campylobacter spp. from different poultry species, then Campylobacter were confirmed by cPCR by detection of 16SrRNA gene, and differentiated by rtPCR into 40 C.jujeni and 62 C.coli by targeting mapA gene amd ceuE gene respectively . After primary isolation of bacteria from Poultry bacterial identification has been investigated using MALDI-TOF Spectrometry, and the following result has obtained. • from poultry cloacal swaps , 28 E. faecium (52.8%), 16 E. faecalis (30.2%) and 9 E. gallinarum (17.0%) strains were obtained . Thirty one Enterococcus isolates isolates were tested for detection of phenotypic antibiotic resistance by using broth microdilution method using MICRONOUT system for Gram positive bacteria and reveled that. • All Enterococcus isolates were multidrug-resistant .All E. faecium isolates were resistant to daptomycin, clindamycin, erythromycin and erythromycin/clindamycin. Four E. faecium isolates were resistant to vancomycin. Resistance rates to other antibiotics ranged between 31.8% for teicoplanin and 86.4% for penicillin G, cefoxitin and fosfomycin. All E. gallinarum isolates showed resistance to cefoxitin, ceftaroline, clindamycin, daptomycin, erythromycin, erythromycin/clindamycin, fosfomycin, fusidic acid, gentamicin and oxacillin. One was resistant to vancomycin. • In 31 phenotypically resistant Enterococcus isolates , The most often identified resistance genes were msrC (n=20) ,erm(B) (n=20). aac6-aph2 and tetL (n=17 each).Vancomycin resistance genes were found in 14 isolates. All E. gallinarum isolates harbored the vanC1 gene but only one of them was phenotypically resistant. (tetK, tetL, tetM, tetS and tetO) were found in 25 isolates. The tetL gene was detected in 18 isolates (13 E. faecium and 5 E. gallinarum) followed by 16 tetM-carrying isolates (12 E. faecium and 4 E. gallinarum). The mecA gene was detected in seven E. faecium and two E. gallinarum by PCR. Sequence analysis of all PCR products obtained in this study were confirmed with the same genes on genBank ,similarity of the obtained sequences and with those on GenBank was 99.0 to 100%. |