Enterobacterales bloodstream infections (E-BSI) cause a significant burden of disease in children and are associated with antimicrobial resistance. We assessed temporal changes in the population-based incidence of E-BSI in children in Queensland, Australia. We conducted a cohort study of incidents of E-BSI occurring in children in Queensland between 2000 and 2019, with a total population of 19.7 million child years. Infections were linked to clinical outcomes in hospital admissions and vital statistics databases. We estimated age- and sex-standardized E-BSI incidence rates over time. Secondary outcomes included the proportion of extended-spectrum β-lactamase phenotypes per year, hospital length of stay, and mortality. We identified 1980 E-BSI in 1795 children. The overall age- and sex-standardized incidence rate was 9.9 cases per 100 000 child years, which increased from 7.3 to 12.9 over the period studied, an increase of 3.9% (95% confidence interval: 3.1-4.7) per year. There were 3.6 cases of E. coli bloodstream infection per 100 000 child years, increasing annually by 4.7% (3.5-5.9). The Salmonella sp. bloodstream infection incidence was 3.0 cases per 100 000 child years, which increased from 2013 by 13.7% (3.8-24.3) per year. The proportion of extended-spectrum β-lactamase E. coli increased over time. Mortality and length of stay were higher among children with comorbidities than those without (4.0% vs 0.3%, and 14 vs 4 days, respectively, P < .001). The age- and sex-standardized incidence of E-BSI almost doubled in Queensland children over 2 decades, driven by increases in Salmonella sp. and E. coli. Increasing resistance of E. coli should prompt the inclusion of children in antimicrobial clinical trials.