Abstract Background & Objectives: The rate of nosocomial infections is a remarkable index that can be used to evaluate the quality and type of health care provided in hospitals. On the other hand, infection of control in hospitals has a direct correlation with the identification of the infectious agents. The aim of this study was to evaluate the level of microbial contamination of operation rooms in Amir-Al-momenin Hospital in Zabol. Materials & Methods: In this cross-sectional descriptive-analytic study, 380 samples were taken from different operating rooms of Hospital. After laboratory analysis, the results were considered with sampling location, sampling time (before or after disinfecting), type of disinfectant and its dilution level. Statistical analysis was done with SPSS soft ware and Pearson chi-square test. Results: The Microbial contamination was 27.6% in the operation rooms of the hospital. Contamination was due to bacterial (18.2%), fungal (7.6%), and bacterial + fungal infections (1.8 %).The most prevalent contaminating agents were Staphylococcus epidermidis (22.5 %), Klebsiella spp (14.2 %) and Aspergillus spp (10.8 %). The most contaminated locations were the floors and walls, the beds and anesthetic masks. The least contaminated area was the air. Conclusion: The microbial contamination rate in Amir-Al-Momenin Hospital of Zabol was not significantly different from other hospitals in the country. It was however, a far cry from world standards (5%). There is a dire need to plan suitable strategies to decrease the microbial contamination rate in the operation rooms of the hospital
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