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This study aimed to determine the bacterial isolates associated with postpartum endometritis among dairy cows in Western Australia and their antimicrobial susceptibility profiles. A cross‐sectional study was conducted between June–October 2020. Endometritis was defined as evidence of mucopurulent to purulent vaginal discharge 60–100 days postpartum. Vaginal discharge samples were obtained, cultured, identified and tested for antimicrobial susceptibility. A total of 118 bacterial isolates were grown from 46 animals, representing 36 species. The bacteria isolated from both aerobic and anaerobic cultures included Bacillus (60.2%), Streptococcus (12.7%), Trueperella (10.1%), Escherichia (6.7%) and Staphylococcus (5.9%). The remaining genera <5% were Histophilus, Aeroccocus, Enterococcus and Moraxella. Resistance was variable between isolates, but the highest resistance levels were observed in Streptococcal and Bacillus isolates to enrofloxacin, clindamycin and erythromycin, respectively. All Streptococcal isolates exhibited 100% resistance to enrofloxacin, and the greatest resistance levels were found in Streptococcus luteinises to trimethoprim‐sulfamethoxazole 83%, clindamycin 66% and 33% quinupristin‐dalfopristin. There was 84.5% resistance to clindamycin and 35.2% to erythromycin in the Bacillus isolates, with the highest resistance found in Bacillus licheniformis and Bacillus subtilis. Escherichia coli exhibited 12.5% resistance to gentamycin, ceftiofur, whereas amoxicillin‐clavulanic acid exhibited 37.5%. Within the Staphylococcal isolates, 28.5%, 28.5%, 42.8% and 14.2% resistance to ceftiofur, erythromycin, cefoxitin, penicillin and tetracycline were observed, respectively. The presence of resistance to important antimicrobials for human use, such as cephalosporins, macrolides and fluoroquinolones, highlights the need for judicious use of antimicrobials in dairy cattle. [ABSTRACT FROM AUTHOR]