Bacterial infection of cow's milk was studied for this aim out of 124 samples of cow's milk were collected from 38 mastitic cow, 46 subclinical mastitis, 10 bulk tank and 30 market milk, 124 these samples were subjected obtained 131 pathogens was detected and the most frequently identified microbes was Staphylococcus aureus 54(43.5٪) followed by Sterptococcus agalacteae 25(20.2٪), Escherichia coli 23(18.5٪), Corynebacterium pyogenes 16(12.9٪), Enterococcus feacalis 10(8.1 ٪) and Mycoplasma Bovis 3(2.4٪). Rate of isolation from different types of milk samples,where 27 isolates where identified from 38 mastitic cow`s milk. S. aureus showed the highest rate 48٪ (number=13), followed by S.agalacteae 26٪(n=7), C. pyogene 19٪(n=5) and lowest persent MB 7٪ (n=2). Concerning subclinical mastitis S. aureus showed the highest rate of isolation 38٪ (n=20), followed by E.coli 28% (n=15), S.agalacteae 19٪ (n=10), C. pyogene 13٪ (n=7) and lowest persent was MB 2٪ (n=1). In as regards to the examined bulk milk, E.coli showed the highest rate of isolation 42%(n=8), followed by S. aureus 37٪ (n=7), C. pyogene 21٪ (n=4) while S.agalacteae and MB were not detected. About the examined marketing milk, S. aureus showed the highest rate 44٪ (n=14), followed by S.agalacteae 25% (n=8) , E.faecalis 31٪ (n=10) while S.agalacteae, E.coli and MB were not detected. Three isolates were identified as MB ( Two isolates from clinical mastitis and one isolate of subclinical mastitis) and confirmed by PCR S.aureus isolates showed multidrug resistance ranged from 60%-100%, where 100% of isolates were resistant to tetracycline, amplicillin, cephalothin, amikacin, clindamycin and lincomycin. S.agalactea showed multidrug resistance ranged from 60%-100%, where 100% of isolates were resistance to tetracycline, neomycin, sulfa/trimethoprim and clindamycin. E.coli showed multidrug resistance ranged from 40%-100%, where 100% of isolates were resistance to sulfa/trimethoprim and lincomycin. C.pyogenes showed multidrug resistance ranged from 61%-100%, where 100% of isolates showed multidrug resistance, and were resistance to tetracycline, amplicillin, neomycin, sulfa/trimethoprim, amikacin and gentamicin. E.faecalis showed multidrug resistance ranged from 20% -100%, where 100% of isolates were resistance to gentamicin and lincomycin. The public health concern of different isolated strains was discussed.