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AbstractEyewash stations are an essential component of laboratory safety programs, providing first aid in case of ocular exposure to hazardous materials. However, the presence of microbial contamination in these devices poses a potential risk of ocular infection to laboratory employees. This cross-sectional study aimed to evaluate the microbial quality and performance of 40 eyewash stations fixed in 10 buildings in a laboratory setting. Water quality parameters, including temperature, pH, turbidity, and the presence of Acanthamoeba spp., were measured at various time points (first draw, after 2 min of flushing, and 15 min flushing) from samples collected from each of the 40 eyewash stations. Performance and operational data were also measured according to the American National Standards Institute (ANSI)/International Safety Equipment Association (ISEA) Z358.1-2014 standard. Our results showed variable compliance with this standard across measures of physical condition, performance, access, and maintenance. Out of the 147 water samples collected (130 eyewash samples, 17 building reference samples), 28 samples were suspected to contain Acanthamoeba spp. or other free-living amoeba based on initial testing. Further analysis using polymerase chain reaction (PCR) confirmed the presence of Acanthamoeba spp. in 5 out of 28 samples. The results of this study provide insights into the potential risk of ocular infections associated with using eyewash stations and provide the basis for the recommendations on maintenance protocols to minimize the risk of microbial contamination. [ABSTRACT FROM AUTHOR]