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 Section
Aspergillus 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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Neighbor-joining 40
88 A. carneus
A. terreus var. africanus

tree of ITS Terrei

95
A. terreus
48 A. allahabadii
87 A. niveus

sequences of 59 Fennellia flavipes

98 A. iizukae Flavipedes
A. carbonarius
A. niger Nigri
selected Aspergilli
91
51 76 A. tubingensis
98 A. campestris
A. candidus Candidi
Candidi
36 Neocarpenteles acanthosporus
50
38 A. clavatus Clavati
• 8 subgenera, 22
Dichotomomyces cejpii
20 95 A. cervinus
52
A. kanagawaensis Cervini
sections 25
47
42
Neosartorya pseudofischeri
A. fumigatus
Neosartorya fennelliae Fumigati
77 A. flavus
99
15
Petromyces alliaceus
A. pseudotamarii Flavi
67 A. auricomus
91
84 A. ochraceus
Circumdati
Neopetromyces muricatus Circumdati
38 A. restrictus Restricti
A. penicillioides Restricti
95 Eurotium echinulatum
98
51 Eurotium rubrum Aspergillus
48 Eurotium chevalieri
30 A. clavatoflavus
5 Zonati
Warcupiella spinosa Warcupi
13 Chaetosartorya chrysella
94 A. wentii Cremei
96 A. rambellii
52 A. ochraceoroseus Ochraceorosei
A. sparsus
95 A. biplanus Sparsi
44 A. raperi Raperi
99 A. ustus
99 A. insuetus
A. pseudodeflectus
Usti
70
74 Emericella rugulosa
Emericella nidulans
96 A. versicolor
64 A. sydowii
Nidulantes
A. protuberus
Sclerocleista thaxteri
99 Sclerocleista ornata Ornati
Monascus purpureus

0.01

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 & Elphick
Section Nidulantes: Emericella Berk. & Br.
Section Flavipedes: Fennellia Wiley & Simmons
Section Circumdati: Neopetromyces Frisvad & Samson
Section Flavi: Petromyces Malloch & Cain
Section 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

3
Conidiophores and conidia

Conidial head

stipe

Footcell

Conidiogenous cells of Aspergillus are phialides

4
 UNISERIATE CONIDIAL HEADS

BISERIATE CONIDIAL HEADS

globose clavate subclavate

SHAPES OF VESICLES IN ASPERGILLUS

5
 Micro-
morphological
criteria
 conidiophores and
conidia
 columnar or radiate
conidial heads
 Hülle cells
 ascomata
 ascospores

RADIATE CONIDIAL HEADS

6
 COLUMNAR CONIDIAL HEADS

COLUMNAR CONIDIAL HEADS

RADIATE CONIDIAL HEADS SPLITTING INTO COLUMNS

7
 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 Hülle

cells of Aspergillus
calidoustus 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 colony Typical columnar heads

Growing fast
Covering the disk
In 5-7 days at 37°C

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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 blue

Drop of alcohol
Preparations should be made from Malt Extract Agar

IDENTIFICATION CRITERIA
 Macro-morphology
Colony color
Colony size
 Micro-morphology
Columnar or radiate conidial heads
Conidial heads: uni- or biseriate
Hülle cells
(Ascomata, ascospores)
 Molecular based identification (sequencing)

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A.calidoustus is also
frequently isolated 65
2gbII27E7IA.ustusIbT
2gbII27D6IA.ustusIbT
2gbII27F3IA.ustusIbT

from indoor 2gbII27E3IA.ustusIbT

2gbII27E2IA.ustusIbT
2gbII27E9IA.ustusIbT

environments gbII27F5IA.ustusIBt
gbII27E8IA.ustusIBt
2gbII27F6IA.ustusIbT A. calidoustus
6 4 2gbII27D3IA.ustusIbT

(partial β-tubulin data)

2gbII27F2IA.ustusIbT
gbII27D8IA.ustusIBt
gbII27E6IA.ustusIBt
gb||CBS112452|N2|AinsuetusCBSB
99 2gbII27D2IA.ustusIbT
gb||20D2|N2|AustusCBSbt2200607
2gbII27F1IA.ustusIbT
51
2gbII27F5IA.ustusIbT
gbII21A2bIA.pseudodeflectusIBt
gbII27E4IA.ustusIBt
89
gb||CBS596.65|N2|AinsuetusCBSB A. pseudodeflectus
99 gb||2CBS107.25|N2|T AinsuetusC
9 7 gb||CBS119.27 - |N2|AinsuetusC A. insuetus
7 7 gbII27E5bIA.ustusIBt
99 gbII27E1IA.ustusIBt Aspergillus sp.
57 gbII27D9IA.ustusIBt
78 gb||CBS561.65|N2|AinsuetusCBSB
98
gb||IBT16751|N2|AgranulosusCBS
37
61
gb||CBS113227|N2|AinsuetusCBSB
gb||CBS209.92|N2|AinsuetusCBSB
A. keveii
gb||CBS119.58|N2|AgranulosusCB
43 gb||2CBS261.67|N2|T AustusCBSB
9 9 gb||20D5 - |N2|AustusCBSbt2200 A. granulosus
gb||27E6|AustusIBT
8 8 gb||CBS495.65|N2|T ApuniceusCB
A. ustus
99 gb||2CBS495.65|N2|T ApuniceusC
7 0 2gbII27D5IA.ustusIbT
96 2gbII27D4IA.ustusIbT A. puniceus
6 5 gb||21A4|N2|AustusCBSBT2200706
gb||27D7I|AustusIbt
44
gb||27E8|AustusIbt
gb||CBS583.65|N2|AversicolorCB
A. versicolor
0.02

88 Aspergillus heteromorphus CBS 117.55 T

100 Aspergillus ellipticus CBS 707.79 T
Aspergillus ellipticus CBS 677.79
Aspergillus carbonarius CBS 101697
Section Nigri
54
Aspergillus carbonarius CBS111.26 NT
93Aspergillus carbonarius CBS 116.49
100Aspergillus carbonarius CBS 126.49 ‘black Aspergilli’
Aspergillus sclerotioniger CBS 115572 T
Aspergillus niger CBS 101883 T
Aspergillus niger CBS 101884
Aspergillus niger CBS 101698
Aspergillus niger CBS 557.65
Aspergillus niger CBS 420.64
100
58 Aspergillus niger CBS 101705
Morphological similar species;
Aspergillus niger CBS 101699
51 Aspergillus niger CBS 101886
Aspergillus niger CBS 618.78
Aspergillus niger CBS 554.65 T
Aspergillus niger CBS 120.49 molecular tools and/or extrolite
Aspergillus brasiliensis CBS 101740
90 Aspergillus vadensis CBS 113365 T
Aspergillus tubingensis CBS 134.48 T
patterns recommended
Aspergillus tubingensis CBS136.52
Aspergillus tubingensis CBS 425.65
8 Aspergillus tubingensis CBS 126.52
Aspergillus tubingensis CBS 161.79
0 Aspergillus tubingensis CBS 115657
Aspergillus tubingensis CBS 117.32
93Aspergillus acidus CBS 564.65 T
85Aspergillus acidus CBS 565.65
53Aspergillus piperis CBS 112811 T
Aspergillus costaricaensis CBS 115574 T
100
Aspergillus homomorphus CBS 101889
92 Aspergillus homomorphus CBS 101888
99 Aspergillus "aculeatus” CBS 620.78
Aspergillus "aculeatus” CBS 114.80
78Aspergillus aculeatus CBS 119.49
Aspergillus aculeatus CBS 172.66 T
100 Aspergillus japonicus CBS 115.80
Aspergillus japonicus CBS 113.48
Aspergillus japonicus CBS 568.65
Aspergillus japonicus CBS 611.78
100
Aspergillus japonicus CBS 101.14
Aspergillus japonicus CBS 522.89
Aspergillus japonicus CBS 114.51 T
Aspergillus flavus CBS 100927 NT
10 changes

www.studiesinmycology.org; www.cbs.knaw.nl;
www.aspergilluspenicillium.org

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