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Surgical risk factors for periprosthetic joint infection (PJI) after total knee arthroplasty (TKA) are the subjects of ongoing research. It is unclear if there are specific locations of the surgical area that might act as a pathogen source. Due to the fact that bacterial replication occurs preferably under humid conditions, it was our aim to investigate if irrigation fluid reservoirs on the surgical covers are subject to bacterial colonization. We prospectively observed 40 patients with scheduled aseptic 1-stage TKA revision. At time intervals of 30 min, irrigation samples were tested for microbiological colonization. Additionally, the suction tip was investigated at the end of surgery. Overall, a bacterial detection rate of 25% was found (57/232 samples). Analysis for any positive microbial detection revealed pathogen findings of irrigation fluid in 41.7% of samples after 30 min with a constant increase up to 77.8% after 90 min. Twenty-three percent of suction tips showed bacterial colonization. Coagulase-negative staphylococci, accounting for the majority of PJI, were the predominant pathogens. After an average follow-up of 17 months, no PJI was confirmed. Despite the substantial bacterial load of irrigation fluid, PJI rates were not elevated. Nevertheless, we recommend that irrigation fluid reservoirs should be prevented and not withdrawn by suction. [ABSTRACT FROM AUTHOR]