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Mould Growth on Building Materials Secondary Matabolites, Mycoxocins and Biomarkers

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Mould Growth on Building Materials Secondary Matabolites, Mycoxocins and Biomarkers Downloaded from orbit.dtu.dk on: Oct 05, 2021 Mould growth on building materials Secondary matabolites, mycoxocins and biomarkers Nielsen, Kristian Fog Publication date: 2001 Document Version Publisher's PDF, also known as Version of record Link back to DTU Orbit Citation (APA): Nielsen, K. F. (2001). Mould growth on building materials: Secondary matabolites, mycoxocins and biomarkers. General rights Copyright and moral rights for the publications made accessible in the public portal are retained by the authors and/or other copyright owners and it is a condition of accessing publications that users recognise and abide by the legal requirements associated with these rights. Users may download and print one copy of any publication from the public portal for the purpose of private study or research. You may not further distribute the material or use it for any profit-making activity or commercial gain You may freely distribute the URL identifying the publication in the public portal If you believe that this document breaches copyright please contact us providing details, and we will remove access to the work immediately and investigate your claim. Mould growth on building materials Secondary metabolites, mycotoxins and biomarkers Kristian Fog Nielsen The Mycology Group Biocentrum-DTU Technical University of Denmark Lyngby 2002 Mould growth on building materials Secondary metabolites, mycotoxins and biomarkers ISBN 87-88584-65-8 © Kristian Fog Nielsen [email protected] Phone + 45 4525 2600 Fax. + 45 4588 4922. The Mycology Group, Biocentrum-DTU Technical University of Denmark, Building 221 Søltofts Plads, Building 221, DK-2800 Kgs. Lyngby, Denmark Energy and Indoor Climate Division Danish Building Research Institute Dr. Neergaards Vej 15, DK-2970 Hørsholm, Denmark Supervisors: Ulf Thrane, associate professor, Ph.D The Mycology Group, Biocentrum-DTU, Technical University of Denmark Suzanne Gravesen, Senior Researcher Energy and Indoor Climate Division, Danish Building Research Institute Examiners: Ib Søndergaard, associate professor, Ph.D Biocentrum-DTU, Technical University of Denmark Brian Flannigan, Professor Napier University, Edinburgh, Scotland Leon Brimer, associate professor, Ph.D Royal Veterinary and Agricultural University Frederiksberg, Denmark This thesis was defended at 14 o’clock the 7. December 2001, in auditorium 51, building 208, Technical University of Denmark. PREFACE More than seven ago years ago I was introduced to the fascinating world of fungi and my- cotoxins by Ole Filtenborg and Thomas O. Larsen. The opportunity to work on my own with an array of sophisticated equipment, made me stay in the Mycology Group, and led to my M.Sc. project where Suzanne Gravesen and Thomas convinced me to work with satratoxins in buildings. Successfully ending this work, it resulted in my Ph.D. project financed by the Danish Building Research Institute where Erik Christophersen organised the funds for the project as well as the many travels. However, it has also been frustrating to work in a field where you are unable to help people, often socially disadvantaged, who are forced to move from their mouldy homes due to mas- sive symptoms, often victims of the ignorance of the authorities. Working in such a multidisciplinary area requires help from a lot of people, and at the Danish Building Research Institute I would like to thank the librarians, especially Lillian Nielsen who have been a tremendous help in ordering scientific papers. Gunnar Holm has been a great help in constructing experimental set-ups, Lotte P. Uttrup and Per Hansen have prepared material samples, Jan Carl Westphall helped with various photographic problems, Yelva Jensen has been a good secretary, and Solveig Nissen with the language revision of some of the papers as well as this thesis. Peter A. Nielsen is acknowledged for the fruitful discus- sions on health and building related issues. Jørgen Ø. Madsen, Department of Organic Chemistry, in preparing the 4-D2-ergosterol was a great help in making the ergosterol analytical method work. The private consultants Mikael Ø. Hansen and Peter Thompson have kindly supplied me with "real" mouldy materials, interesting field observations, and taken their time to show me the real mouldy world outside the laboratories. I would like to thank Dr. Aino Nevalainen, Dr. Maija-Riitta Hirvonen, and their helpful Ph.D. students for letting me work in their laboratories at the National Public Health Institute, Ku- opio, Finland. Working at TNO Bouw, The Netherlands, with Dr. Olaf Adan and Gerben van der Wel helped me understand the importance of instationary environmental conditions for mould growth. Having Professor Bruce B. Jarvis in our group for a year raised the level of my work on Stachybotrys and natural products significantly and introduced me to a number of other re- search groups. The help of colleagues in the Mycology group, in particular Elisabeth Krøger, Kir Lyhne, Flemming Lund, Thomas O. Larsen, Jørn Smedsgaard, Jens C. Frisvad and Birgitte Ander- sen has been encouraging. Ulf Thrane has been the perfect Ph.D. supervisor who always had time - if needed, and on the other hand never intervened much in my work. Thanks to Suzanne Gravesen who believed so much in my work and have taught me so much about so many different things and introduced me to so many people as well as help- ing with the language revision of this report. The support and understanding of my beloved wife Hanne - especially during my stays in Finland, Holland and various conference as well as the acceptance of the large piles of pa- pers, CD, books etc. during my project cannot be overestimated. Lyngby, June 2001 Kristian Fog Nielsen Page i ABSTRACT The aim of this study was to document if the moulds produce mycotoxins and other biologi- cally active metabolites when growing in buildings, as well as investigate the influence of environmental conditions on the production of these metabolites. The growth of moulds un- der various humidities should also be investigated along with the use of chemical biomarkers for quantitation of mould growth. It was shown that Stachybotrys chartarum produced a number of mycotoxins when growing in buildings. These components were produced in significantly higher quantities than by other moulds investigated in this study. Only 35% of the isolates from buildings produced the ex- tremely cytotoxic satratoxins. Actually these metabolites are probably not responsible for idiopathic pulmonary hemosiderosis in infants, which is probably caused by other S. charta- rum metabolites. For the first time ever Stachybotrys metabolites were found in air-samples, where several classes of spiriocyclic drimanes and satratoxins were detected. Aspergillus versicolor produced high quantities of the carcinogenic mycotoxin, sterigmato- cystin at water activities (aw) > 0.95. At lower aw more than 10 unknown metabolites were produced, including at least 5 metabolites also produced by A. ochraceus. A. versicolor was often growing in mixed cultures with others moulds where it sporulated poorly, meaning that it may evade detection based on cultivating methods. The A. ustus isolates from buildings were macro-morphologically and chemically very different from the cereal isolates, and should be described as a new species. Penicillium chrysogenum produced few detectable metabolites and often none when growing on materials. Combined with the no observed effects on persons experimentally exposed to high quantities of the spores, these observations implies that this species may not be impor- tant and is actually obscuring the detection of more toxic genera and species. P. brevicom- pactum produced mycophenolic acid and P. polonicum the tremorgenic verrucosidin when they were inoculated on water-damaged materials. Chaetomium globosum produced high quantities of chaetoglobosins whereas Trichoderma species did not produce detectable quantities of trichothecenes when growing on materials. Even on laboratory media <1% of the isolates produced trichodermol or esters of it. Ergosterol content of building materials was quickly and precisely quantified by isotope dilu- tion GC-MS/MS. Determination of ergosterol is only needed as a supplement for assessing mould growth on test materials, as visual assessment, especially supported by dissection microscopy generally was just as sensitive. The minimal RH for growth on wood based mate- rials and material containing starch was just below 80% at room temperature, and increased to about 90% at 5°C. On paper-mineral composites such as gypsumboard the minimal RH was approx. 90% RH from room temperature to 5°C. Pure mineral based materials with few organic additives seem to be able to support growth at RH ≥ 0.90, although ≥95% RH was needed to generate chemical detectable quantities of biomass. The phylloplane Cladosporium was able to outgrow P. chrysogenum on materials under transient humidities. This is presumably why phylloplanes like Cladosporium, Ulocladium, Phoma and Aureobasidium are very common in bathrooms and other places with instation- ary humidity conditions. Mould growth in buildings is causing various health effects among the occupants, however the causal components is still partly unknown making scientifically based guidelines for "how much is too much" and cost efficient remediation of mouldy buildings almost impossible. Page ii RESUME Målet med dette studium var at dokumentere om skimmelsvampe producerer mykotoksiner og andre biologisk aktive stoffer, når de vokser på byggematerialer.
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