A total number of 228 apparently healthy milk samples were collected from individual and bulk tank milk of cows (100 and 86 samples, respectively), goats (30) and she camel (12) for isolation of some lactic acid bacteria (LAB) especially that have coccal form. The preliminary screening LAB community at the genus level depending on the basis of morphological characteristics showed that, the isolates were differentiated into 4 groups; Enterococci, Leuconostocs, Pediococci and Streptococci with a total percentage of 61%. The highest % of LAB was recorded for Enterococcus species in the different animal species especially in camel milk (41.7%). Antibacterial activity of selected 75 LAB strains against S. aureus, S. uberis, E. coli and Yersinia enterocolotica as bovine mastitis pathogens were detected. 53 out of 75 of the selected strains showed antibacterial effect against the tested pathogens. Eighteen Enterococcus isolates have inhibitory effects on all of the tested bacteria with inhibition zone diameter ranged between 10-25 mm. Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) was used as an aid step for identification of LAB strains. Thus the SDS-PAGE results confirmed the biochemical identification of the isolated cultures for Leuconostoc mesenteroides with a percentage of similarity (90.8%), for Pediococcus acidilactici (92.5%), for Enterococcus hirae (99.84%) and for Streptococcus thermophilus (99.89%). Representative strains of genus Enterococci that had higher antibacterial activity against mastitis pathogens were subjected to sequence-based identification. The obtained sequences of these isolates were submitted to the Gen Bank database with accession numbers KU847974 and KU847975 for E. faecium and E. hirae, respectively and showed 99% 16S rRNA sequence homology. It was concluded that raw animal milk may be a po­tential source for the isolation of probiotic LAB with antibacterial properties against mastitis pathogens that may be presented as an interesting alternative to antibiotic drugs to overcome the antibiotic resistance of mastitis pathogens as well as antibiotic residues in milk.