The study aimed to investigate the phenotypic and genotypic identification of in vitro antimicrobial susceptibility of 21 Staphylococci (10 Staphylococcus aureus and 11 Coagulase Negative Staphylococci) isolated from bovine mastitis to 12 antimicrobial drugs frequently using in veterinary medicine in Algeria. Isolates of staphylococci from bovine mastitis were tested for antibiotics with disc-diffusion method according to the National Committee for Clinical Laboratory Standards guidelines in the Mueller-Hinton agar, and resistant genes mecA, blaZ, aac-aph, ermA, ermC, tetK and tetM were detected by PCR. Staphylococci isolates showed high resistance to penicillin (95.23%), oxacillin (80.95%), clindamycine (80.95%), and erythromycin (76.19%) but, no resistance in all these strains was detected for gentamicin. Among 21 isolates of Staphylococci, 20 were found to be methicillin and multidrug resistant. Multidrug resistant strains exhibited several antibiogram patterns (antibiotic I to XIII). The distribution of antibiotic-resistant genes was mecA (100%) and tetM (100) followed by blaZ (42.85%). In the present study, the significant determination was the high prevalence of methicillin-resistant Staphylococci, which were resistant to multiple antibiotics. The finding of methicillin-resistant staphylococci from bovine mastitis is the first report in Algeria and revealed the status of resistant isolates in herd that might be helpful in treatment, controlling of resistant strains and for deciding culling of cows.