Summary: | (1) Background: As β-lactamase-producing Enterobacterales are no longer exclusively associated with the health care system, investigating the potential risk they pose to the integrity of the environment and food safety has become of utmost importance. This study aimed to determine the prevalence of extended-spectrum β-lactamase (ESBL), AmpC, and carbapenemase-producing Enterobacterales isolates from retailed raw vegetables and to determine if household washing is an effective method of lowering bacterial load; (2) Methods: Seasonal vegetables (<i>n</i> = 165) were acquired from supermarkets (<i>n</i> = 2) and farmer markets (<i>n</i> = 2) in Romania. Following sample processing and isolation, identification of Enterobacterales was performed by matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF). Polymerase chain reaction (PCR) multiplex was used to ascertain the presence of the main ESBL, AmpC, and Carbapenemase genes. Phenotypic antibiotic resistance profiles of isolates were determined by extended antibiograms. <i>Enterobacteriaceae</i> colony-forming units (CFU) counts were compared between vegetable types; (3) Results: Beta-lactamase producing bacteria were observed on 7.9% of vegetables, with 5.5% displaying ESBL/AmpC phenotype and 2.4% identified as Carbapenemase producers. The most frequently detected β-lactamase genes were <i>bla</i><sub>SHV</sub> (<i>n</i> = 4), followed by <i>bla</i><sub>CTX-M</sub> and <i>bla</i><sub>TEM</sub> (each with <i>n</i> = 3). Phenotypic antibiotic resistance analysis showed that 46% of isolates were multiple drug resistant, with aminoglycosides (38.5%) the most prevalent non-β-lactam resistance, followed by first-generation quinolones (38.5%). (4) Conclusions: The present study has described for the first time the presence of β-lactamase producing Enterobacterales in fresh produce retailed in Romania.
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