Background: Antimicrobial-resistant bacteria originating from vegetable farms represent a growing global concern with serious public health implications. Continuous assessment, surveillance, and monitoring of antimicrobial resistance along the farm-to-consumer pathway are therefore essential. This study aimed to assess the antimicrobial susceptibility profiles of selected bacterial species belonging to the Gammaproteobacteria, isolated from irrigation water, soil, farms, and supermarket vegetables. Methods: A cross-sectional study was conducted in central Ethiopia from February to June 2024. A total of 408 samples (soil, irrigation water, vegetable from farms and supermarkets) were collected from six formal vegetable farms and supermarkets in central Ethiopia. Identification was performed using conventional biochemical tests and matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS). All isolates were tested for their susceptibility to 16 antimicrobials following Clinical laboratory standards institute (CLSI) guidelines using Kirby Bauer disk diffusion assay. Chi-square test was used to assess difference of antimicrobial resistance among bacteria isolates between sampling types, with a p -value of less than 0.05 considered statistically significant. A multivariable binary logistic regression analysis was employed to identify factors associated with multidrug resistance (MDR), with sampling types and genus. Results: Out of the total (331) isolates, the dominant species identified were Escherichia coli and Enterobacter spp., followed by Citrobacter spp. The proportion of bacterial genera was highest in vegetable samples from farms (32.3%), followed by vegetables obtained from supermarkets (18.6%), soil (16.4%), and irrigation water (14.2%). The highest resistance rate (40%) was observed in E. coli isolates from supermarket vegetables to ampicillin and piperacillin/tazobactam, while highest rate of resistance to ceftazidime was observed among Enterobacter spp. from irrigation water (37.5%). Overall, 33.2% of isolates were susceptible to all antimicrobials, 24.7% were resistant to at least one antibiotic, and 20.8% were multidrug resistant (MDR). MDR occurrence was highest in vegetables isolates (26.4%), followed by soil (19.4%) and irrigation water (19%). The highest MDR prevalence was observed in Klebsiella oxytoca (60%), followed by Klebsiella aerogenes (33.3%). Conclusion: The current study highlights the presence of multidrug resistant environmental bacteria, including Enterobacteriaceae family and Acinetobacter spp. throughout vegetable production from farms to supermarkets. The presence of MDR environmental isolated Enterobacteriaceae family and Acinetobacter spp. throughout vegetable production and supermarkets emphasizes the importance of increased surveillance of antimicrobial resistance (AMR) in fresh produce and the production environment within a One Health paradigm to develop AMR control strategies.

Occurrence of antimicrobial resistance (AMR) and antimicrobial resistance genes (ARGs) in Enterobacteriaceae and Acinetobacter spp. within agricultural environments represents a growing global health concern. Vegetables have been increasingly recognized as reservoirs of multidrug‐resistant (MDR) pathogens. The study aimed to detect ARGs in Enterobacteriaceae and Acinetobacter spp. from vegetable market chains and assess the clonal relationship of isolates. Antimicrobial susceptibility testing was conducted for a total of 170 isolates obtained from vegetables, irrigation water, and soil samples, and ARGs were detected by polymerase chain reaction (PCR). Conjugation assay was conducted, and genetic relatedness among bacterial isolates was assessed by ERIC‐PCR at a 70% similarity cut‐off. High resistance rate was observed to sulfamethoxazole/trimethoprim (34.0%) and cefepime (28.8%). Phenotypically, 23.5% (40/170) of isolates were extended spectrum β ‐lactamase (ESBL) producers and from meropenem resistance isolates 60% (6/10) were carbapenemase producers. Genes encoding for broad‐spectrum β ‐lactamases were detected in 30% (12/40) tested isolates, including those from irrigation water (50%; 3/6), vegetables (30.7%; 8/26) and soil (12.5%; 1/8) samples. Carbapenemase genes were detected in carbapenem resistant isolates from irrigation water (80%; 4/5) and vegetable (40%; 2/5) samples. The most prevalent β ‐lactamase genes identified were bla CTX‐M and AmpC ( n = 5 each), bla SHV ( n = 2), and bla TEM ( n = 1). Among carbapenemase genes bla NDM ( n = 6) was frequently detected, and bla KPC and bla VIM in a single isolate each. A sul1 gene was identified in 33.3% of isolates from vegetable and 15.3% of those from irrigation water. Four transconjugants were detected and successfully transferred resistance genes with conjugation frequency ranging from 1.48 × 10 ⁻⁵ to 3.3 × 10 ⁻⁶ out of 15 tested. ERIC‐PCR revealed diverse clonal lineages, although K. pneumoniae , K. aerogenes and E. coli isolates from vegetables and irrigation water displayed close genetic similarity (100%, 70%, and 78%), respectively. This study provided the first evidence of diverse β ‐lactamases and carbapenemase‐producing Enterobacteriaceae in Ethiopian vegetable market chains. These findings highlight the need for continued surveillance and improved food safety measures to mitigate potential public health risks posed by antimicrobial‐resistant pathogens in the food supply chain.