The use of antimicrobials in livestock production and their effect on the development of antimicrobial resistance (AMR) is a global health problem for humans, animals and the environment. The aim of this study was to determine antimicrobial residue levels in milk and feces samples during the withdrawal period in dairy cattle administrated with a single dose of the drug, as well as to characterize the antimicrobial resistance patterns of Escherichia coli cultured from feces samples. In the study, dairy cows from three different farms in North Macedonia were included. Raw milk and feces samples were collected before drug administration (0 day) and on the 1st, 2nd, 3rd, 7th and 21st day after drug administration. The antimicrobial residues of oxytetracycline, enrofloxacin, amoxicillin, trimethoprim and procaine-benzylpenicillin were determined using a validated liquid chromatography combined with tandem mass spectrometry (LC-MS/MS) method involving stable isotopes. According to results obtained, the highest levels of analyzed antimicrobial residues were determined on the first day after drug administration, which then gradually decreased until their elimination (7th day). The highest AMR of E. coli (100%) was found in β-lactam antimicrobials. Less exposure to broad-spectrum antimicrobials could be an important factor for reduction of AMR on dairy farms.

Commensal Escherichia coli has the potential to easily acquire resistance to a broad range of antimicrobials, making it a reservoir for its transfer to other microorganisms, including pathogens. The aim of this study was to determine the prevalence of resistant commensal Escherichia coli isolated from dairy cows’ feces. Phenotypic resistance profiles and categorization were determined by minimum inhibitory concentration (MIC) testing with the broth microdilution method, while the PCR method was used to determine the presence of resistant genes. Out of 159 commensal E. coli isolates, 39 (24.5%) were confirmed to have resistance. According to the MIC values, 37 (97.3%) and 1 (2.7%) isolate were phenotypically categorized as ESBL and ESBL/AmpC, respectively. All isolates showed resistance to ampicillin, while 97.4%, 56.4%, and 36% showed resistance to cefotaxime, ciprofloxacine, and azitromycine, respectively. Not all isolates that showed phenotypic resistance were found to be carrying the corresponding gene. The most prevalent resistant genes were gyrA, tetA, sul2, and tetB, which were present in 61.5%, 64%, 54%, and 49% of the isolates, respectively. The results clearly indicate that, besides their resistance to multiple antimicrobials, the commensal E. coli isolates did not necessarily carry any genes conferring resistance to that particular antimicrobial.