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Background: Despite the high risk of bacterial contamination, no studies have been found that evaluate the optimal hanging time of open-system enteral sets under room temperature and intensive care conditions. This study aims to determine the differences in microbial loads of solutions taken from open-system enteral feeding sets hanging for 24 h under room temperature and intensive care conditions., Methods: This study is a descriptive type of research. The study sample consisted of a total of 40 samples, with 20 samples under room temperature (25.2-23.3°C) and 20 samples under intensive care conditions (21.1-24.2°C). A simple random sampling method was used for sample selection. The assessment of microbial quality was based on the US Food and Drug Administration online Bacteriological Analytical Manual., Results: In this study, it was determined that 5% of the samples taken from intensive care and 15% of the samples taken at room temperature exceeded the accepted limit for aerobic mesophilic bacteria count. The count of catalase and coagulase-negative Staphylococcus spp was 5% in intensive care samples and 40% in room temperature samples. The total coliform count was not detected in intensive care samples but was 35% in room temperature samples. Escherichia coli was not detected in either group., Conclusion: Differences in microbial loads were observed when commercial enteral solutions were administered using open-system enteral feeding sets under room and intensive care conditions for 24 h. Therefore, large-scale studies are needed to determine the safe use of open-system enteral feeding sets under room temperature conditions., (© 2024 American Society for Parenteral and Enteral Nutrition.)