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To clarify the clinical and microbial characteristics of polymicrobial bacteremia (PM to contribute to improvements in clinical diagnosis and effective early treatment. This retrospective multicenter study used data from three acute-care hospitals in Okayama Prefecture, Japan, collected between January 2014 and March 2019. We reviewed the demographics, comorbidities, organisms isolated, infectious focus, and 30-day mortality of patients with PMB. Of the 7233 positive blood cultures, 808 (11.2%) were positive for more than one organism.