Mastitis in cows is a major problem in dairy farms leading to a decrease in the quantity and quality of milk. The aim of the present study was to examine the association between the presence of Escherichia coli (E. coli) in milk and the subclinical mastitis, and to characterize the antibiotic resistance profiles of the isolated E. coli. In the current study, a total of 360 cow raw milk samples from three dairy farms of the region of Algiers were analyzed. The analysis period lasted from Spring 2017 to Winter 2019. The California Mastitis Test (CMT) was applied to detect subclinical mastitis. The E. coli strains were isolated from milk using conventional bacteriological methods. The antibiotic resistance profile of the isolated E. coli strains to 12 different antibiotics was tested using the disk diffusion method. On β-lactamase-producing strains, a double diffusion test was applied to identify the Extendedspectrum β-lactamase (ESBL) phenotype. Finally, the ctXx-M genes were amplified by PCR. Two-thirds (66.4%) of the milk samples were positive for the CMT test. A total of 97 E. coli strains were isolated from the milk samples, their resistance to antibiotics was tested, and 3.1% of the strains were resistant to trimethoprim-sulfamethoxazole, 6.2% to chloramphenicol, 12.3% to gentamicin, 13.4% to colistin, 23.3% to amoxicillin/clavulanate, 31.9% to kanamycin, 39.2% to enrofloxacin, 51.5% to cefotaxime, 52% to tetracycline, 57.7% to ampicillin, 74.3% to nalidixic acid, and 75.3% to amoxicillin. Furthermore, most of the E. coli strains (92.8%) were resistant to more than one antibiotic with a Multiple Antibiotic Resistance index ranging from 0 to 0.8. The 50 strains resistant to cefotaxime were analyzed for an ESBL phenotype. 39 of them (78%) were positive to the double-disk synergy test. Among the 39 ESBL positive strains, 27 (69.2%) were confirmed for the presence of a CTX-M gene by PCR. The present study showed that multiple drug-resistant E. coli, including ESBL-carriers, were frequently isolated from the milk of dairy cows in Algeria. The results underlined that the use of antibiotics on farms must be reasoned to avoid the spread of resistant strains in animals and human populations.