Background: Bloodstream infections (BSIs) are one of the most frequently observed hospital-acquired infections (HAIs). Objectives: We aimed to describe the epidemiology and drug resistance of hospital-acquired Enterobacteriaceae BSIs and to check for any correlation with the type of hospital care. Methods: In 2015–2018, 333 Enterobacteriaceae isolates were collected from hospitalized internal medicine and surgical patients. The drug-resistance testing was conducted according to the EUCAST recommendations, using the disc-diffusion method to determine resistance to penicillin, cephalosporins, carbapenems, aminoglycosides, fluoroquinolones, and sulfamethoxazole with trimethoprim. Tests confirming the presence of extended-spectrum β-lactamases (ESBLs) and KPC, NDM, and OXA-48 carbapenemases were performed. We determined the minimum inhibitory concentration (MIC) values (mg/L) for selected antibiotics. To detect the resistance genes, a single PCR reaction, a multiplex PCR, and a real-time PCR were conducted. Results: The prevalence rate of Enterobacteriaceae bacilli in BSIs was 23.5%. Penicillin resistance remained at a very high level of almost 100%, with only the piperacillin-tazobactam resistance remaining at 19%–22%. The same was true for cephalosporins: the bacilli have only shown a high susceptibility to cefoperazone with sulbactam (4%–14% of them were resistant). Ciprofloxacin (53%–62%) and sulfamethoxazole with trimethoprim (48–55%) have proven highly resistant. Carbapenems were the only antibiotics with susceptibility at 98%–99%. No difference was found between the types of hospital care (surgical vs nonsurgical) and the levels of antimicrobial resistance in the studied Enterobacteriaceae isolates (Table 1). Conclusions: The high prevalence of Enterobacteriaceae bacilli in BSI is particularly worrying, as is the high rate of resistance to cephalosporins and aminoglycosides, which are often used in the empirical therapy. Unfortunately, our results indicate the need to base the empirical therapy on carbapenems.Funding: This work was supported by a grant from Jagiellonian Univerity Medical School (No. N41/DBS/000053)Disclosures: None