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To investigate microorganisms causing central venous catheter contamination and how this contamination differs across different catheter metrics.After obtaining IRB approval and informed consent, 830 cultures were prospectively obtained from 45 ICU patients with central venous catheter or peripherally inserted central catheter. Bacterial colonies were identified by mass spectrometry.Bacterial contamination of central catheter hubs occurred 44% of the time in this study in the ICU setting. Coagulase-positive staphylococci cultures had higher median (± interquartile range) CFUs (12 ± 232) versus coagulase-negative (3 ± 10) and other bacteria (1 ± 3; P < 0.001). Bacterial contamination was associated with various metrics. Higher incidence (P < 0.05) of coagulase-positive staphylococci cultures was associated with hub-only connections (a “hub” being a female connection; 10.9% vs. 7.9% male connections), connections without a manifold (1 lumen device that mixes multiple infusions together; 9.7% vs. 0% with manifold); and central venous pressure monitoring connections (25.8% vs. 7.1% without). Internal jugular sites (10.0% vs. 2.7% femoral, 6.2% PICC, P = 0.031) and medial lumens of triple lumen catheters (11.9% vs. 5.6% distal, 7.0% proximal, P = 0.049) had increased incidence of higher bacteria loads (> 15 CFUs).This study found a high incidence of central access catheter hub bacterial contamination, which correlated with positive blood cultures in 2 of 3 total bacteremia cases identified in the 45 patients.Central-line associated blood stream infections increase length of hospital stay and costs of care.The needleless connectors (that is “hubs”) of the central line catheters in the intensive care unit had a high incidence (44%) of bacterial contamination.Further studies are needed to examine if bacterial contamination of the needless connectors may help identify patients at risk for central-line associated blood stream infections.