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CBS-KNAW Fungal Biodiversity Centre, UtrechtCBS-KNAW Fungal Biodiversity Centre, Utrecht
Course
FOOD and AIRBORNE FUNGI
Course leaders and assistants
Rob Samson Jos Houbraken Tineke van Doorn Martin Meijer Neriman Yilmaz
Telephone, fax and Email
Telephone 030 2122600 Telefax 030 2512097
CBS has a wireless network for laptops (ask password)
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8-12 October 2012 Programme Monday October 8
10.00 Opening10.10 Introduction (R.A. Samson)11.30 Zygomycetes
12.00-15.00 Microscopy Zygomycetes 15.00 Aspergillus 15.20 -17.00 Microscopy Aspergillus
Tuesday October 910.00 Methods for isolation of food-borne Fungi (J. Houbraken)11.00 Ascomycetes
11.30-15.00. Microscopy Ascomycetes15.00 Penicillium15.30–17.00 Microscopy Penicillium
Wednesday October 1010.00 Molecular methods for identifying food-borne fungi. An overview (J. Houbraken)11.00 Microscopy Penicillium13.30 Fusarium14.30–17.00 Microscopy Fusarium
15.00 Conidiogenesis of Deuteromycetes
Thursday October 1110.00 Detection of Airborne fungi (R.A. Samson)11.30–17.00 Microscopy Deuteromycetes
Friday October 12
10.00 Mycotoxins 10.45 Yeasts 11.00 -14.00 continuation Microscopy Deuteromycetes 14.00 General DiscussionP
rogramme of the course
Dinner together with the course participants
3 course menu is Euro 19.75For some dishes there is an extra charge
Course from 9.00-17.00 Lecture mostly starting 10.00
and 15.30 Coffee and tea breaks Lunch 13.00-13.30
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Lunch 13.00
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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)
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Pro
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urse 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
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Baarn
Utrecht
AmsterdamAmsterdam
Rotterdam
CBS was founded in 1904and is since 1968 an institute ofthe Royal Netherlands Academyof Arts and Sciences. The institute moved in December 2000 from Baarn to Utrecht
Frits Went (1863-1935) professor in botany at the
University of Utrecht
The initiator of CBS
Prof. Johanna Westerdijk(1883-1961)
Phytopathologist andthe first female professorin 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 65.000 strains are preserved
for long-term maintenance
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• 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)
Research programmes
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Penicillium roqueforti on rye bread
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Bathroom
Pathogenic Fungi
There are relatively very few fungi which are dangerous for man and animalsHowever 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
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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
Aflatoxins•Aspergillus flavus•Aspergillus parasiticus•Aspergillus nomius
MYCOTOXINS
Immune surpression
Tremorgenic activityCarcinogenic
Gastro-intestinal disturbances
Nephrotoxic
Parasympathetic nerve system
Hepatic necrosis
Oestrogenic
Haemorrhage of thelung and brain
Haemaorrhageof liver
Dermal toxicosis
Idiopathic pulmonaryhemosiderosis
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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 indoorenvironments 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
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Growth of fungi
Source Carbon Temperature 0-60°C, optimal 25-
30°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 structuresSaccharomyces cerevisiae, a yeast with one celled structures
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Penicillium expansum in culture
Apples contaminated with Penicillium expansum
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Mycelium of a Mucor
hyphahypha
hyphahypha
SporangiophoreSporangiophore
Coenocytic mycelium of a ZygomyceteCoenocytic mycelium of a Zygomycete
Mycelium of a hyphomycete (Wallemia sebi)Mycelium of a hyphomycete (Wallemia sebi)
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Coniophora puteana hyphe with clamp connections
Numbers of known and estimated total species of organisms
vascular plants 220.000 270.000bryophytes 17.000 25.000algae 40.000 60.000fungi 75.000 >1.500.000bacteria 3.000 >1.000.000viruses 5.000 130.000
group known taxa total taxa
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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
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Rhizopus stolonifer
rhizoids
stolon
Rhizopus
apophysis
columella
Sexual production in the Zygomycetes
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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
Ascoma (cleistothecium)
Ascus with ascospores
Sexual reproduction in the Ascomycetes
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Ascospores of Eurotium herbariorum
Emericella nidulans
Anamorph: Aspergillus nidulans
Hülle cells
Deuteromycetes or anamorphic fungi
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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
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Blastic conidiogensis in Botrytis
Poroconidia in Alternaria
Phialides with conidia in chains
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The common mycoflora in food and in indoor environments
Zygomycetes (about 10-15 species) Ascomycetes (about 10-15 species) Deuteromycetes (about 100-150
species)
Absidia corymbiferaAcremonium mumorumAcremonium strictumAlternaria alternariaAspergillus candidusAspergillus clavatusAspergillus flavusAspergillus flavipesAspergillus fumigatusAspergillus nigerAspergillus ochraceusAspergillus penicillioidesAspergillus restrictusAspergillus sydowii Aspergillus terreusAspergillus versicolorAureobasidium pullulansBotrytis cinereaCandida peltataChaetomium aureumChaetomium globosumChaetomium indicum Chrysonillia sitophilaCladosporium cladosporioidesCladosporium herbarumCladosporium sphaerospermumClonostachys roseaCoprinus cordisporusCryptococcus laurentii
Curvularia lunataEmericella nidulansEpicoccum nigrumEurotium amstelodamiEurotium chevalieriEurotium herbariorumExophalia dermatitidisFusarium culmorumFusarium solaniFusarium verticillioidesGeomyces pannorumGeotrichum candidumMemnoniella echinataMucor hiemalisMucor plumbeusMucor racemosusOidiodendron griseumOidiodendron rhodogenumPaecilomyces lilacinusPaecilomyces variotiiPenicillium aurantiogriseumPenicillium brevicompactumPenicillium chrysogenumPenicillium citrinumPenicillium communePenicillium corylophilumPenicillium expansumPenicillium glabrumPenicillium janthinellium
Penicillium olsoniiPenicillium rugulosumPenicillium simplicissimumPenicillium spinulosumPenicillium variabilePhialophora fastigiataPhialophora verrucosaPhoma glomerata Phoma macrostomaPithomyces chartarumPyronema domesticumRhizopus stoloniferRhodotorula mucilaginosaSchizophyllum communeScopulariopsis brevicaulisScopulariopsis candidaScopulariopsis fuscaSerpula lacrymansSistotrema brinkmaniiSporobolomyces roseusStachybotrys chartarumSyncephalastrum racemosumTrichoderma harzianumTrichoderma koningiiTrichoderma virideTritirachium oryzaeUlocladium chartarumVerticillium lecaniiWallemia sebi
List of fungal species occurring in indoor environments
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)
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0.02
Beta tubulin phylogeny of section Fumigati
N. hiratsukae CBS 294.93
N. multiplicata CBS 646.95
A. unilateralis CBS 126.56
A. turcosus KACC 42091
N. assulata KACC 41691
N. tatenoi CBS 407.93
N. pseudofischeri CBS 208.92
N. nishimurae IFM 54133
A. clavatus CBS 513.65
N. aurata CBS 466.65N. stramenia CBS 498.65
N. galapagensis CBS 117522
N. fennelliae CBS 598.74N. denticulata CBS 652.73a
N. ferenczii NRRL 4179N. glabra CBS 111.55
N. udagawae CBS 114217N. aureola CBS 105.55
A. viridinutans CBS 127.56
A. novofumigatus IBT 16806A. fumigatiaffinis IBT 12703
A. fumisynnematus IFM 42277A. lentulus FH5
A. fumigatus CBS 133.61N. fischeri CBS 544.65
N. spinosa CBS 483.65A. laciniosa KACC 41657
N. coreana KACC 41659
A. duricaulis CBS 481.65A. brevipes CBS 118.53
N. quadricincta CBS 135.52
N. spathulata CBS 408.89
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N. papuensis CBS 841.96
N. australensis CBS 112.55
N. warcupii NRRL 35723
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Introduction to the Zygomycetes Microscopical examination of
cultures
