Background The epidemiology of enterococcal bacteraemia has changed worldwide and vancomycin-resistant enterococci increasingly cause healthcare-associated infections) with limited treatment options. Studies show heterogeneity among countries, regions and individual hospitals. Methods We retrospectively analysed enterococcal bacteraemia with Enterococcus faecalis and E. faecium at the largest hospital in Croatia, University Hospital Centre Zagreb from January 2017 to December 2021. Results A total of 432 cases of enterococcal bacteraemia were identified with 256 (59.3%) due to E. faecalis and 176 (40.7%) to E. faecium. Enterococcal bacteraemia occurred more frequently in men (n = 270; 62.5%) and the median age of all patients was 62 years (IQR: 0–92). We found statistically significant increase in the incidence trend of bacteraemic episodes with an annual percent change of 20.9% (95% confidence interval 14.3 to 27.8; p = .002) predominantly due to an increase of E. faecalis bacteraemia. The majority of patients (362/432; 83.8%) had healthcare-associated infections and 38.0% (165/432) of patients were in the intensive care unit. The proportion of vancomycin-resistant enterococcal bacteraemia increased from 12.7% (n = 8/63) in 2017 to 25.7% (n = 29/113) in 2021, statistically significant increasing trend (p = .0455), mainly due to an increased proportion of vancomycin-resistant E. faecium (p = .0169). Conclusions This is the first study describing the trends in enterococcal bacteraemia and vancomycin-resistance in enterococci in Croatia. We found a rising trend in enterococcal bacteraemia and in the proportion of vancomycin resistance and identified the most vulnerable patient groups, notably intensive care unit patients.