Detection of viable fungal spores contaminant on documents and rapid control of the effectiveness of an ethylene oxide disinfection using ATP assay

• Published in: Luminescence (Chichester, England) Vol. 18; no. 2; pp. 113 - 121
• Main Authors: Rakotonirainy, Malalanirina S., Héraud, Cécile, Lavédrine, Bertrand
• Format: Journal Article
• Language: English
• Published: Chichester, UK John Wiley & Sons, Ltd 01.03.2003
• Subjects: Adenosine Triphosphate - analysis; Algorithms; Aspergillus - chemistry; Aspergillus niger - chemistry; ATP bioluminescence; ATP extraction; biodeterioration; Disinfectants; Disinfection; Ethylene Oxide; fungi; Hot Temperature; Indicators and Reagents; Libraries; rapid microbiology; Reference Standards; Reproducibility of Results; Species Specificity; Spores, Fungal - chemistry; Sterilization
• ISSN: 1522-7235; 1522-7243
• DOI: 10.1002/bio.710

Filamentous fungi are able to damage and even destroy archival and library materials. Nowadays the conventional method for detecting such micro‐organisms is to put them in cultures but such methods are laborious and time‐consuming. ATP methodology has been widely applied in other domains and its success on bacteria and yeast has been demonstrated. Several commercial reagent kits are available but they did not give satisfactory results on spores mould. We have elaborated new extraction strategies specific to fungi. A comparison of 42 extraction protocols of ATP from fungal spores was carried out. Extraction at 100°C with DMSO 90% in a Tris–acetate–EDTA buffer proved to be the best method. The viability of cells is estimated by the determination of adenylate energy charge (EC). We applied our method successfully on well‐known species such as Aspergillus flavus, A. niger, A. fumigatus, A. versicolor, Neosartorya fischeri, Eurotium chevalieri, Penicillium chrysogenum, Chaetomium globosum and Ulocladium spp. The results suggest that the ATP bioluminescence assay provides a sensitive and time‐saving method for detecting viable fungal spores. The validity of the procedure was also tested on spores killed by steam and on spores treated with ethylene oxide. We showed that EC determination could be used for a rapid control of the effectiveness of a disinfection process performed with ethylene oxide. Copyright © 2003 John Wiley & Sons, Ltd.