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This study showed a strong association between building renovation and an increase in environmental Aspergillus contamination. Results confirmed the high efficacy of laminar airflow plus HEPA filtration and a high air-change rate. Although filtration with HEPA was effective during normal conditions, it alone was unable to prevent the rise of Aspergillus contamination related to building renovation. This study emphasized the necessity of an environmental survey of airborne contamination related to construction, to facilitate prevention of nosocomial aspergillosis outbreaks. A standardized protocol for aerobiological surveillance is needed.