Programme of the course. Telephone, fax and 04/10/2012. Course. FOOD and AIRBORNE FUNGI. Course leaders and assistants

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1 Course FOOD and AIRBORNE FUNGI CBS-KNAW Fungal Biodiversity Centre, Utrecht Course leaders and assistants Rob Samson Jos Houbraken Tineke van Doorn Martin Meijer Neriman Yilmaz Programme of the course Telephone, fax and Telephone Telefax CBS has a wireless network for laptops (ask password) 1

2 Programme of the course 8-12 October 2012 Programme Monday October Opening Introduction (R.A. Samson) Zygomycetes Microscopy Zygomycetes Aspergillus Microscopy Aspergillus Tuesday October Methods for isolation of food-borne Fungi (J. Houbraken) Ascomycetes Microscopy Ascomycetes Penicillium Microscopy Penicillium Wednesday October Molecular methods for identifying food-borne fungi. An overview (J. Houbraken) Microscopy Penicillium Fusarium Microscopy Fusarium Conidiogenesis of Deuteromycetes Thursday October Detection of Airborne fungi (R.A. Samson) Microscopy Deuteromycetes Friday October Mycotoxins Yeasts continuation Microscopy Deuteromycetes General Discussion Dinner together with the course participants 3 course menu is Euro For some dishes there is an extra charge Programme of the course Course from Lecture mostly starting and Coffee and tea breaks Lunch

3 Programme of the course Lunch Programme of the course CBS rules Please wear name badges all the time No smoking or food in the laboratory Please wear laboratory coat when possible NO mobile phones in the course room Use only one strain each time and return the plates Do not leave the plates open to avoid release of spores in the air Be careful with the burners and put them off when you are not using it Try to work with your neighbour(s) Programme of the course Introduction to the fungal flora of food and the indoor environment Introduction to the important groups of Zygomycetes, Ascomycetes and Deuteromycetes Microscopical examination of cultures Lectures on applied aspects 3

4 Rotterdam Amsterdam Baarn Utrecht CBS was founded in 1904 and is since 1968 an institute of the Royal Netherlands Academy of Arts and Sciences. The institute moved in December 2000 from Baarn to Utrecht Frits Went ( ) professor in botany at the University of Utrecht The initiator of CBS Prof. Johanna Westerdijk ( ) Phytopathologist and the first female professor in the Netherlands Centraalbureau voor Schimmelcultures the oldest and largest culture collection of filamentous fungi and yeasts Since 2001 also including the NCCB bacteria and phages collections more than strains are preserved for long-term maintenance 4

5 Research programmes Applied and Industrial Mycology (R.A. Samson) Bioinformatics group (V. Robert) Fungal Physiology (R. de Vries) Origin of Pathogenicity in Clinical Fungi (G.S. de Hoog) Collection, Preservation and Digitalisation (G. Verkleij) Evolutionary Phytopathology (P.W. Crous) Yeast and Basidiomycete Research (T. Boekhout) 5

6 Penicillium roqueforti on rye bread 6

7 Bathroom Pathogenic Fungi There are relatively very few fungi which are dangerous for man and animals However when the immune system is affected opportunistic species which can commonly occur become pathogenic Aspergillus fumigatus mostly not common in indoorenvironments important in clinical environments when present, it can be easily be detected in air samples incubation temperature important quantitative data should be treated carefully 7

8 Penicilllium chrysogenum An important and useful mould because of the production of the antibiotic penicillin Very common species on food and in indoor environments can produce several mycotoxins Aflatoxins Aspergillus flavus Aspergillus parasiticus Aspergillus nomius Tremorgenic activity Haemorrhage of the lung and brain Immune surpression Carcinogenic Nephrotoxic Oestrogenic Hepatic necrosis Haemaorrhage of liver MYCOTOXINS Gastro-intestinal disturbances Idiopathic pulmonary hemosiderosis Dermal toxicosis Parasympathetic nerve system 8

9 Stachybotrys chartarum common in temperate and humid climates produces mycotoxins (e.g. satratoxins) respiratory tract bleeding non-respiratory symptoms Indication to cause death in infants Mycotoxins in the indoor environments Fungi which occur in indoor environments are similar to those found in food These fungi are also able to produce mycotoxins These toxic metabolites can be inhaled directly or via the spores Where are the Fungi? In principle everywhere on earth 9

10 Growth of fungi Source Carbon Temperature 0-60 C, optimal C ph 1-12, optimal at neutral ph water oxygen The biodiversity of fungi on food and in indoorenvironments Saccharomyces cerevisiae, a yeast with one celled structures 10

11 Penicillium expansum in culture Apples contaminated with Penicillium expansum 11

12 hypha hypha Sporangiophore Mycelium of a Mucor Coenocytic mycelium of a Zygomycete Mycelium of a hyphomycete (Wallemia sebi) 12

13 Coniophora puteana hyphe with clamp connections Numbers of known and estimated total species of organisms group known taxa total taxa vascular plants bryophytes algae fungi > bacteria > viruses

14 Zygomycetes: Mucor, Rhizopus, Absidia Ascomycetes: Eurotium, Emericella, Neosartorya Basidiomycetes: Agaricus, Serpula Deuteromycetes belong either to the Ascomycetes or Basidiomycetes but they do not have a sexual form (teleomorph): Penicillium, Aspergillus, Cladosporium Yeasts are Ascomycetes or Basidiomycetes Zygomycota or Zygomycetes 14

15 rhizoids stolon Rhizopus stolonifer Rhizopus columella apophysis Sexual production in the Zygomycetes 15

16 Ascomycota; Ascomycetes Diverse group of fungi, multiplying in a yeast-like manner without fruit-bodies or produce mycelium with fruit-bodies (= ascomata) Worldwide distribution; soil, dung, plants, food and feed Saprobes or parasites to humans, animals, plants In food or indoor environments a limited number of genera occur and are mostly very simple. Eurotium: ascoma and asci Ascus with ascospores Ascoma (cleistothecium) Sexual reproduction in the Ascomycetes 16

17 Ascospores of Eurotium herbariorum Emericella nidulans Anamorph: Aspergillus nidulans Hülle cells Deuteromycetes or anamorphic fungi 17

18 Conidiogenesis Conidiogenesis = the mode of conidium formation Conidia = specialized non-motile asexual spore Conidiogenous cell = specialized cell which give rise to the conidia Conidiophore = entire system of fertile hyphae Thallic and blastic conidiogenesis Thallic conidia in Geotrichum candidum 18

19 Blastic conidiogensis in Botrytis Poroconidia in Alternaria Phialides with conidia in chains 19

20 The common mycoflora in food and in indoor environments Zygomycetes (about species) Ascomycetes (about species) Deuteromycetes (about species) List of fungal species occurring in indoor environments Curvularia lunata Absidia corymbifera Emericella nidulans Penicillium olsonii Acremonium mumorum Epicoccum nigrum Penicillium rugulosum Acremonium strictum Eurotium amstelodami Penicillium simplicissimum Alternaria alternaria Eurotium chevalieri Penicillium spinulosum Aspergillus candidus Eurotium herbariorum Penicillium variabile Aspergillus clavatus Exophalia dermatitidis Phialophora fastigiata Aspergillus flavus Fusarium culmorum Phialophora verrucosa Aspergillus flavipes Fusarium solani Phoma glomerata Aspergillus fumigatus Fusarium verticillioides Phoma macrostoma Aspergillus niger Geomyces pannorum Pithomyces chartarum Aspergillus ochraceus Geotrichum candidum Pyronema domesticum Aspergillus penicillioides Memnoniella echinata Rhizopus stolonifer Aspergillus restrictus Mucor hiemalis Rhodotorula mucilaginosa Aspergillus sydowii Mucor plumbeus Schizophyllum commune Aspergillus terreus Mucor racemosus Scopulariopsis brevicaulis Aspergillus versicolor Oidiodendron griseum Scopulariopsis candida Aureobasidium pullulans Oidiodendron rhodogenum Scopulariopsis fusca Botrytis cinerea Paecilomyces lilacinus Serpula lacrymans Candida peltata Paecilomyces variotii Sistotrema brinkmanii Chaetomium aureum Penicillium Sporobolomyces roseus Chaetomium globosum aurantiogriseum Stachybotrys chartarum Chaetomium indicum Penicillium Syncephalastrum racemosum Chrysonillia sitophila brevicompactum Trichoderma harzianum Cladosporium cladosporioides Penicillium chrysogenum Trichoderma koningii Cladosporium herbarum Penicillium citrinum Trichoderma viride Cladosporium Penicillium commune Tritirachium oryzae sphaerospermum Penicillium corylophilum Ulocladium chartarum Clonostachys rosea Penicillium expansum Verticillium lecanii Coprinus cordisporus Penicillium glabrum Wallemia sebi Cryptococcus laurentii Penicillium janthinellium Toxinogenic pathogenic xerophilic bath rooms and wet environments Identification of moulds There are presently no fast methods for identification of genus or species Morphological examination is essential New molecular approaches are developed The new morphological data show that some common species are species complexes Examples are Fusarium, Cladosporium, Aspergillus In Penicillium and Aspergillus a polyphasic approach is used, combining morphology, biochemistry and molecular data (for example β- tubulin and calmodulin sequences) 20

21 Beta tubulin phylogeny of section Fumigati 100 N. warcupii NRRL A. unilateralis CBS N. papuensis CBS N. assulata KACC N. aurata CBS N. stramenia CBS N. galapagensis CBS N. multiplicata CBS N. australensis CBS N. fennelliae CBS N. denticulata CBS a N. ferenczii NRRL 4179 N. glabra CBS N. nishimurae IFM A. turcosus KACC N. tatenoi CBS N. aureola CBS N. udagawae CBS A. viridinutans CBS A. laciniosa KACC N. spinosa CBS A. fumigatiaffinis IBT A. novofumigatus IBT N. coreana KACC N. fischeri CBS A. fumigatus CBS A. lentulus FH5 A. fumisynnematus IFM N. hiratsukae CBS N. spathulata CBS N. pseudofischeri CBS N. quadricincta CBS A. brevipes CBS A. duricaulis CBS A. clavatus CBS Programme of the course Introduction to the Zygomycetes Microscopical examination of cultures 21