1
Identification of Aspergillus
Aspergillus a common genus with
approx. 300 species several ascomycetous
teleomorphs has preference for
warmer climates important for
biotechnology, producers of mycotoxins and opportunistic human pathogens
Nomenclature of infrageneric taxa of the genus Aspergillus (group-names following the classification of Raper and Fennell,1965)
Subgenus SectionAspergillus Aspergillus (A. glaucus-group)
Restricti (A. restrictus-group)Fumigati Fumigati (A. fumigatus-group)
Cervini (A. cervinus-group)Ornati Ornati (A. ornatus-group)Clavati Clavati (A. clavatus-group)Nidulantes Nidulantes (A. nidulans-group)
Versicolor (A. versicolor-group)Usti (A. ustus-group)Terrei (A. terreus-group)Flavipedes (A. flavipes-group)
Circumdati Wentii (A. wentii-group)Flavi (A. flavus-group)Nigri (A. niger-group)Circumdati (A. ochraceus-group)Candidi (A. candidus-group)Cremei (A. cremeus-group)Sparsi (A. sparsus-group)
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99
99
70
64
96
74
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7799
98
84
4088
96
95
87
95
94
91
76
52
91
44
98
30
48
67
59
13
38
5
5148
15
20 95
51
38
4247
3650
52
25
0.01
Monascus purpureusSclerocleista ornataSclerocleista thaxteri
A. raperiA. ustusA. insuetusA. pseudodeflectus
Emericella rugulosaEmericella nidulans
A. versicolorA. sydowiiA. protuberus
A. rambelliiA. ochraceoroseusA. sparsusA. biplanus
Eurotium echinulatumEurotium rubrum
Eurotium chevalieriA. clavatoflavus
Warcupiella spinosaChaetosartorya chrysellaA. wentii
A. restrictusA. penicillioides
A. auricomusNeopetromyces muricatusA. ochraceus
A. flavusPetromyces alliaceus
A. pseudotamarii
A. cervinusA. kanagawaensis
Neosartorya pseudofischeriA. fumigatus
Neosartorya fennelliae
A. campestrisA. candidus
Neocarpenteles acanthosporusA. clavatus
Dichotomomyces cejpii
A. carbonariusA. nigerA. tubingensis
A. carneusA. terreus var. africanusA. terreus
A. allahabadiiA. niveus
Fennellia flavipesA. iizukae
Nidulantes
Ornati
Usti
Raperi
Ochraceorosei
Sparsi
CremeiWarcupi
Zonati
Aspergillus
Restricti
Circumdati
Flavi
Fumigati
Cervini
Clavati
Flavipedes
Nigri
Candidi
TerreiNeighbor-joining tree of ITS sequences of selected Aspergilli
• 8 subgenera, 22 sections
CircumdatiRestricti
Candidi
Ascomycetous genera with an Aspergillus anamorph
Aspergillus anamorph Teleomorph
Section Aspergillus: Eurotium Link:Fr. Section Fumigati: Neosartorya Malloch & Cain. Section Ornati: Warcupiella Subram., Sclerocleista Subram.,
Hemicarpenteles Sarbhoy & ElphickSection Nidulantes: Emericella Berk. & Br. Section Flavipedes: Fennellia Wiley & SimmonsSection Circumdati: Neopetromyces Frisvad & SamsonSection Flavi: Petromyces Malloch & CainSection Cremei: Chaetosartorya Subram.
Overview identification criteria
Macro-morphology:Colony color
Colony size
Micro-morphology
Molecular based identification(sequencing)
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Colony colors and size in Aspergillus
Colony colors and size in Aspergillus
Micro-morphological criteria
conidiophores and conidia
columnar or radiate conidial heads
Hülle cells
ascomata
ascospores
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Conidiophores and conidia
Footcell
stipe
Conidial head
Conidiogenous cells of Aspergillus are phialides
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UNISERIATE CONIDIAL HEADS
BISERIATE CONIDIAL HEADS
SHAPES OF VESICLES IN ASPERGILLUS
clavateglobose subclavate
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Micro-morphological criteria
conidiophores and conidia
columnar or radiate conidial heads
Hülle cells
ascomata
ascospores
RADIATE CONIDIAL HEADS
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COLUMNAR CONIDIAL HEADS
COLUMNAR CONIDIAL HEADS
RADIATE CONIDIAL HEADS SPLITTING INTO COLUMNS
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Micro-morphological criteria
columnar or radiate conidial heads
conidiophores and conidia
Hülle cells
ascomata
ascospores
Emericella nidulans Anamorph: Aspergillus nidulans
Hülle cells
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Hülle cells
conidia
Irregularly shaped Hüllecells of Aspergilluscalidoustus embedded in conidia
Micro-morphological criteria
columnar or radiate conidial heads
conidiophores and conidia
Hülle cells
ascomata
ascospores
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Example:Aspergillus fumigatus
mostly not common in indoor environments
important in clinical environments
incubation temperature important
Blue green colonyGrowing fastCovering the diskIn 5-7 days at 37°C
Typical columnar heads
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Typical isolates Clinical isolates
Aspergillus penicillioides(xerophilic, no growth on MEA and CYA)
Aspergillus versicolor (strong odour; moderate xerophilic)
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MEDIA FOR IDENTIFICATION
Malt Extract Agar (MEA) for macro- and micro morphology
Czapek Yeast Agar (CYA) for colony color and size
For xerophilic Aspergilli: MEA and CZ with 20% sucrose
Incubation temperature: 25°C Incubation time: 7 days (or longer) 3 point position
Microscopic preparation of Aspergillus
Lactic acid with analine blue/cotton blueDrop of alcoholPreparations should be made from Malt Extract Agar
IDENTIFICATION CRITERIA
Macro-morphologyColony color
Colony size
Micro-morphologyColumnar or radiate conidial heads
Conidial heads: uni- or biseriate
Hülle cells
(Ascomata, ascospores)
Molecular based identification (sequencing)
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2gbII27E7IA.ustusIbT 2gbII27D6IA.ustusIbT
2gbII27F3IA.ustusIbT 2gbII27E3IA.ustusIbT 2gbII27E2IA.ustusIbT 2gbII27E9IA.ustusIbT
gbII27F5IA.ustusIBt gbII27E8IA.ustusIBt 2gbII27F6IA.ustusIbT 2gbII27D3IA.ustusIbT 2gbII27F2IA.ustusIbT gbII27D8IA.ustusIBt
gbII27E6IA.ustusIBt gb||CBS112452|N2|AinsuetusCBSB 2gbII27D2IA.ustusIbT gb||20D2|N2|AustusCBSbt2200607 2gbII27F1IA.ustusIbT 2gbII27F5IA.ustusIbT
gbII21A2bIA.pseudodeflectusIBt gbII27E4IA.ustusIBt gb||CBS596.65|N2|AinsuetusCBSB
gb||2CBS107.25|N2|T AinsuetusC gb||CBS119.27 - |N2|AinsuetusC
gbII27E5bIA.ustusIBt gbII27E1IA.ustusIBt
gbII27D9IA.ustusIBt gb||CBS561.65|N2|AinsuetusCBSB gb||IBT16751|N2|AgranulosusCBS
gb||CBS113227|N2|AinsuetusCBSB gb||CBS209.92|N2|AinsuetusCBSB
gb||CBS119.58|N2|AgranulosusCB gb||2CBS261.67|N2|T AustusCBSB
gb||20D5 - |N2|AustusCBSbt2200 gb||27E6|AustusIBT
gb||CBS495.65|N2|T ApuniceusCB gb||2CBS495.65|N2|T ApuniceusC 2gbII27D5IA.ustusIbT 2gbII27D4IA.ustusIbT
gb||21A4|N2|AustusCBSBT2200706 gb||27D7I|AustusIbt gb||27E8|AustusIbt
gb||CBS583.65|N2|AversicolorCB
4399
88
70
44
65
96
99
57
7799
6137
98
78
9799
51
89
99
65
64
0.02
A. ustus
A. puniceus
A. granulosus
A. keveii
A. versicolor
A. calidoustus
A. pseudodeflectus
A. insuetusAspergillus sp.
A.calidoustus is also frequently isolated from indoor environments
(partial β-tubulin data)
Aspergillus heteromorphus CBS 117.55 TAspergillus ellipticus CBS 707.79 TAspergillus ellipticus CBS 677.79
Aspergillus carbonarius CBS 101697Aspergillus carbonarius CBS111.26 NTAspergillus carbonarius CBS 116.49
Aspergillus carbonarius CBS 126.49Aspergillus sclerotioniger CBS 115572 TAspergillus niger CBS 101883 TAspergillus niger CBS 101884Aspergillus niger CBS 101698Aspergillus niger CBS 557.65Aspergillus niger CBS 420.64Aspergillus niger CBS 101705Aspergillus niger CBS 101699Aspergillus niger CBS 101886Aspergillus niger CBS 618.78Aspergillus niger CBS 554.65 TAspergillus niger CBS 120.49Aspergillus brasiliensis CBS 101740
Aspergillus vadensis CBS 113365 TAspergillus tubingensis CBS 134.48 TAspergillus tubingensis CBS136.52Aspergillus tubingensis CBS 425.65Aspergillus tubingensis CBS 126.52Aspergillus tubingensis CBS 161.79Aspergillus tubingensis CBS 115657Aspergillus tubingensis CBS 117.32Aspergillus acidus CBS 564.65 TAspergillus acidus CBS 565.65Aspergillus piperis CBS 112811 TAspergillus costaricaensis CBS 115574 T
Aspergillus homomorphus CBS 101889Aspergillus homomorphus CBS 101888
Aspergillus "aculeatus” CBS 620.78Aspergillus "aculeatus” CBS 114.80
Aspergillus aculeatus CBS 119.49Aspergillus aculeatus CBS 172.66 TAspergillus japonicus CBS 115.80Aspergillus japonicus CBS 113.48Aspergillus japonicus CBS 568.65Aspergillus japonicus CBS 611.78Aspergillus japonicus CBS 101.14 Aspergillus japonicus CBS 522.89Aspergillus japonicus CBS 114.51 T
Aspergillus flavus CBS 100927 NT10 changes
10088
58
100
90
80
51
100
9354
538593
92100
99
100
78
100
Section Nigri‘black Aspergilli’
Morphological similar species;molecular tools and/or extrolite patterns recommended
www.studiesinmycology.org; www.cbs.knaw.nl; www.aspergilluspenicillium.org