Visual session of unusual : images and plates

J. Varga, S. Kocsubé, G. Szigeti, N. Baranyi, J. Houbraken, R. A. Samson Aspergillus

Aspergillus terreus Pietro Antonio Micheli (Nova conidial head Aspergillum (holy Plantarum Genera, 1729) water sprinkler) Outline of the presentation

• Overview of Aspergillus & nomenclature

• Unusual/newly described Aspergilli in the clinical setting – Section Fumigati – Section Flavi – Section Nigri – Section Nidulantes – Section Circumdati – Other species

Aspergillus taxonomy and nomenclature • What is Aspergillus? What is Aspergillus?

“Fennellia monodii” “Aspergillus paradoxus”

“Penicillium inflatum” Phylogenetic Nidulantes Raperi Usti relationships within the Usti

Aspergillus genus Sparsi Terrei Usti Flavipedes Candidi • type strains of 92 species were Nigri examined Flavi

• 6 genes: Acl1, MCM7, RPB1, RPB2, Circumdati Tsr1, Cct8 • maximum likelihood (PhyML) Fumigati Clavati Cervini Cremei Use of dual nomenclature in Aspergilli • Teleomorph: Neosartorya fischeri (Wehmer) Malloch & Cain 1972 • Anamorph: Aspergillus fischerianus Samson & W. Gams 1985

One = One name

Aspergillus is largely a monophyletic genus Possible name if the */* CBS 272.89 Aspergillus togoensis */* CBS 553.77T Aspergillus coremiiformis */99 NRRL 3357 Aspergillus flavus genus is kept as a */* CBS 151.66T Aspergillus leporis NT 0.96/- CBS 109.46 Aspergillus avenaceus */* CBS 108.08NT Aspergillus ochraceus unit CBS 112812T Aspergillus steynii */99 CBS 649.93T Aspergillus robustus 0.94/- */* CBS 260.73T Fennellia flavipes subgenus Circumdati 1.00/- CBS 653.74T Aspergillus aureofulgens Incl. sections Circumdati, Flavi, 1.00/93 NIH 2624 Aspergillus terreus Aspergillus 0.96/- CBS 118.45T Aspergillus janus Candidi, Flavipedes, Nigri, */99 CBS 566.65NT Aspergillus candidus Terrei CBS 463.65NT Aspergillus arenarius -/- */* CBS 172.66T Aspergillus aculeatus CBS 513.88 Aspergillus niger */- */* CBS 245.65 Aspergillus versicolor */99 CBS 264.81 Aspergillus sydowii 0.98/51 FGSC A4 Emericella nidulans */* CBS 600.67T Aspergillus amylovorus subgenus Nidulantes NT */96 */98 CBS 656.73 Aspergillus egyptiacus Aspergillus CBS 121611 Aspergillus calidoustus Incl. sections Nidulantes, Usti, */82 CBS 707.71T Aspergillus bisporus Ochraceorosei, Sparsi, (Aeni) */96 CBS 101887 Aspergillus ochraceoroseus */59 */* CBS 116.56T Aspergillus funiculosus */90 CBS 139.61NT Aspergillus sparsus CBS 468.65NT Aspergillus biplanus */85 CBS 476.65NT Aspergillus conjunctus */* NRRL 1NT Aspergillus clavatus */90 CBS 558.71T Neocarpenteles acanthosporus */* CBS 157.66NT Dichotomomyces cejpii subgenus Fumigati Aspergillus */* NT */96 NRRL 181 Neosartorya fischeri Incl. sections Fumigati, Clavati Af293 Aspergillus fumigatus */* NT and Cervini */51 CBS 538.65 Aspergillus kanagawaensis CBS 196.64NT Aspergillus cervinus */* CBS 109945T Phialosimplex chlamydosporus */* CBS 128032T Phialosimplex canicus */* CBS 380.74T Basipetospora halophilica */* CBS 384.61T Polypaecilum insolitum Phialosimplex & Polypaecilum Aspergillus */* CBS 101166 Polypaecilum pisci -/- CBS 366.77T Phialosimplex sclerotialis */89 CBS 578.65NT Aspergillus pulvinus */94 CBS 525.83T Cristaspora arxii */97 CBS 104.07NT Aspergillus section Cremei Aspergillus CBS 127.61NT Aspergillus brunneo-uniseriatus */64 */* CBS 516.65NT Eurotium herbariorum */77 CBS 518.65NT Eurotium amstelodami */* subgenus Aspergillus CBS 117.33NT Aspergillus restrictus DTO 11C3 Aspergillus penicillioides Incl. sections Restricti, Aspergillus Aspergillus Wisconsin 54-1255 Penicillium chrysogenum SIM 70 (2011) 1-51 Distribution of Aspergillus sections based on sequence-based identification in the transplant associated infection surveillance network

Usti Nidulantes Nigri Terrei 3% 3% 9% 5%

Flavi 13% Fumigati 67%

J Clin Microbiol 2009; 47: 3138-41; 3271-75 Polyphasic DNA (ITS, -tubulin, calmodulin, Actin, TEF) taxonomic concept Morphology (colony & micromorphology) for Aspergilli

Physiology (effect of temperature/pH, C-source utilization, CREA) Species

Extrolites (Ehrlich test, HPLC-DAD)

Ecology (host, geographical origin) Genetics Genomics (mating tests, VCG) (comparative genomics) Species with Species with known genome sequencing genome sequences in progress

• Aspergillus fumigatus (3 isolates) • Aspergillus tubingensis • Aspergillus clavatus • Aspergillus welwitschiae • Aspergillus brasiliensis • Aspergillus flavus • Aspergillus kawachii (?) • Aspergillus nidulans • Aspergillus luchuensis • Aspergillus niger (3 isolates) • Aspergillus sojae • Aspergillus oryzae • Aspergillus versicolor • Aspergillus terreus • Aspergillus sydowii • Neosartorya fischeri • Aspergillus wentii • Aspergillus carbonarius • Aspergillus glaucus • Aspergillus aculeatus • Aspergillus zonatus • Aspergillus parasiticus • Aspergillus cervinus • Aspergillus sparsus • Aspergillus tubingensis • Aspergillus penicillioides

Aspergillus section Fumigati Aspergillus fumigatus Fresenius 1863

• A. fumigatus is the most frequent cause of IA in most medical centers • People at risk are transplant recipients, patients with HIV/AIDS, with chronic granulomatous disease, with severe combined immunodeficiency, etc. • ca. 350 000 patients suffer from IA worldwide according to Fungal Research Trust (2011) • Able to grow at/above 50°C

Fresenius’s original drawing of A. fumigatus conidial heads Aspergillus fumigatus

CYA MEA

Fast-growing blue- green Typical columnar heads colonies covering the entire MEA plate in 5-7 days at 37°C Aspergillus fumigatus

Typical isolates Conidiophores in sputum Clinical isolates Synonyms of A. fumigatus described from clinical material • A. aviarius Peck 1891 • A. bronchialis Blumentritt 1901 • A. septatus Sartory & Sartory 1943 • A. fumigatus var. ellipticus Raper & Fennell 1965 • A. fumigatus var. minimus Sartory 1919 • A. phialiseptatus Kwon-Chung 1975 • A. neoellipticus Kozakiewicz 1989 • A. acolumnaris Kozakiewicz 1989 • A. arvii Aho et al. 1994 • A. syncephalis Gueguen 1904 • A. lignieresi Cost. & Lucet 1905 Tree of N. warcupii NRRL 35723 A. unilateralis CBS 126.56 N. papuensis CBS 841.96 Aspergillus N. assulata KACC 41691 100 N. aurata CBS 466.65 N. stramenia CBS 498.65 section Fumigati N. galapagensis CBS 117522 N. multiplicata CBS 646.95 N. australensis CBS 112.55  N. fennelliae CBS 598.74 based on -tubulin 86 100 N. denticulata CBS 652.73a N. ferenczii NRRL 4179 sequence data N. glabra CBS 111.55 N. nishimurae IFM 54133 A. turcosus KACC 42091 N. tatenoi CBS 407.93 97 N. aureola CBS 105.55 N. udagawae CBS 114217 A. viridinutans CBS 127.56 95 A. laciniosa KACC 41657 N. spinosa CBS 483.65

76 98 A. fumigatiaffinis IBT 12703 A. novofumigatus IBT 16806 N. coreana KACC 41659 86 N. fischeri CBS 544.65 85 A. fumigatus CBS 133.61 99 A. lentulus FH5 A. fumisynnematus IFM 42277 N. hiratsukae CBS 294.93 N. spathulata CBS 408.89 N. pseudofischeri CBS 208.92 N. quadricincta CBS 135.52 A. brevipes CBS 118.53 A. duricaulis CBS 481.65 100 A. clavatus CBS 513.65

0.02 Stud Mycol 2007; 59: 147-203 Species identification

• Anamorphs can hardly be distinguished by morphological means • ITS can be used to define “species complexes” (e.g. A. fumigatus complex, A. niger complex) • If more precise identification is needed, the use sequences of parts of other genes (e.g. calmodulin, -tubulin)

A. fumigatus A. lentulus A. fumisynnematus A. fumigatiaffinis Distribution of section Fumigati strains not growing at 48°C in Spain

A. viridinutans A. thermomutatus 14% 7%

A. fumigatiaffinis 7% A. lentulus 50% A. fumisynnematus 4%

A. hiratsukae 18%

Antimibrob Agents Chemother 2008; 52: 1244-1251

Clinical presentation of feline SOA

SOA clinical signs: • exophthalmos • corneal ulceration • oral mass / ulcer • nasal signs variably present • paranasal soft tissue swelling • neurological signs 15% cases Dr.Dr. M.M. DowdenDowden

Dr. J. Angles Dr. E. Koelmeyer

35

Pathogenesis - SOA

• Progression from SNA, an invasive mycosis

• Lysis of orbital bone most common site for spread into orbit Aspergillus felis sp. nov. An emerging pathogen in cats, dogs and humans

V. Barrs1, T. van Doorn2, P. Martin1, J. Houbraken2,S. Kidd3, M. Richardson, D. Pinheiro, J. Varga, R. A. Samson2.

1Valentine Charlton Cat Centre, University of Sydney 2CBS-KNAW Fungal Biodiversity Centre, Utrecht 3Mycology Unit, W & C Hospital, Adelaide, Australia Aspergillus felis sp. nov.

Barrs et al PLOS One 2013 2 μm A. felis - host range & disease

• Dogs (n=1):

•9 y MN Old English Sheepdog systemic aspergillosis

•IMHA – prednisolone, cyclosporin

• Humans (n=5):

•A cause of chronic IPA - refractory to Rx (ITZ, POS,VCZ, CSF).

•Phylogenetic evidence – 4 other human isolates:

Invasive pulmonary aspergillosis (Spain) Sputum (Spain) Nail (Spain) Clinical specimen (Japan) MAT-1 MAT-2

cleistothecia

anamorph complementary mating types teleomorph Heterothallic fungus

100 μm

cleistothecium 8-spored asci ascospore

82 131F9 131G2 131F2

84 131E6 Aspergillus felis sp. nov. 131F6 131G3 131E5 88 99 131G1 A. fumigatus var. sclerotiorum CBS 458.75T 87 98 “A. viridinutans” NIHAV2 Aspergillus sp. nov. 90 “A. viridinutans” IMI 182127

99 N. udagawae CBS 114217 99 N. aureola CBS 105.55 A. viridinutans CBS 127.56 A. viridinutans KACC 41142 100 A. viridinutans IMI 367415 N. spinosa CBS 483.65 A. fumigatiaffinis IBT 12703 A. novofumigatus IBT 16806

96 A. lentulus FH5 91 131F5 A. fumisynnematus IFM 42277 N. fischeri CBS 544.65 A. fumigatus CBS 133.61 131E8 100 131F7 131F8

N. fennelliae CBS 598.74 N. hiratsukae CBS 294.93 N. pseudofischeri CBS 208.92 99 131G4 A. clavatus CBS 513.65

0.05

Emerg Infect Dis 2009; 15: 1292-1294 A. wyomingensis

• Identified from an 18 month old British Short Haired cat came from Melbourne in Australia (V. Barrs, personal communication)

Aspergillus section Flavi

O O

Aflatoxin B1 O

O O OCH3 Aspergillus flavus

• Usually considered as the 2nd most prevalent cause of IA • Frequently encountered in keratitis cases and as causative agent of ABPA • Limited molecular data indicate that most clinical isolates belong to A. flavus, with one assigned to A. alliaceus

Diagn Microb Infect Dis 2003; 46: 25-28

J Clin Microbiol 2007; 45: 3464-3467

A. nomius

Aspergillus pseudotamarii has recently also been identified in a keratitis case in India

Aspergillus section Nigri

HOOC OH O O

NH O

Ochratoxin A Cl

ochratoxin A Species distribution of black Aspergilli from various studies

Balajee et al. 2009 Alcazar-Fuoli et al. 2009

A. acidus A. tubingensis 9% 32% A. niger 38%

A. niger A. tubingensis 68% 53%

Hong et al. 2009 Howard et al. 2011

unknown A. niger A. niger 7% 13% 30% A. tubingensis 18%

A. tubingensis A. awamori A. acidus 70% 55% 7% Species assignment of ca. 200 black Aspergilli came from clinical environment Species distribution of 200 black Aspergilli

A. A. aculeatus aculeatinus 0.5% A. awamori 0.5% 16.1% A. acidus 6.5% A. tubingensis A. niger 58.8% 17.6%

Preliminary results of antifungal susceptibility tests indicate species-specific differences (in progress) A. tubingensis from keratitis cases

Cornea 2009; 28:951-4 A. brasiliensis from keratitis cases in India

• The species was described in 2007 by our group • Occurs in soil (Brazil, Australia, USA, Netherlands), on grapes (Portugal), and in human keratitis (India) Black Aspergilli in otomycosis cases

Species distribution of black Aspergilli from otomycosis cases in Iran and Hungary

A. welwitshciae Iran

A. tubingensis A. niger

Hungary A. tubingensis

A. welwitschiae 100 A. neoniger CBS 115656 A. neoniger CBS 115657 Tree based on A. eucalypticola CBS 122712 A. vadensis CBS 113365 A. costaricaensis CBS 115574 calmodulin A. piperis CBS 112811 95 74 A. luchuensis CBS 564.65 72 A. coreanus CBS 119384 sequence data A. tubingensis CBS 134.48 A. niger clade

99 A. brasiliensis CBS 101740 A. welwitschiae CBS 557.65 A. niger CBS 554.65 100 A. foetidus CBS 114.49 97 83 A. foetidus CBS 121.28 A. lacticoffeatus CBS 101883 A. carbonarius CBS 111.26 80 95 A. sclerotioniger CBS 115572 A. sclerotiicarbonarius CBS 121057 A. carbonarius clade A. ibericus CBS 121593

A. ellipticus CBS 707.79 A. heteromorphus clade 100 A. heteromorphus CBS 117.55 A. homomorphus CBS 101889 A. homomorphus clade 97 A. fijiensis CBS 313.89 A. fijiensis CBS 119.49 100 96 A. aculeatinus CBS 1212060 A. aculeatus CBS 172.66

99 A. uvarum CBS 121591 A. indologenus CBS 114.80 A. aculeatus clade A. japonicus CBS 114.51 87 A. violaceofuscus mut. grisea CBS 122.35 99 A. violaceofuscus CBS 123.27

76 A. violaceofuscus CBS 102.23 A. bahamensis CBS 115571 A. flavus CBS 100927 Stud Mycol 69 (2011) 1-17 20

A. neoniger (identified in 2 keratitis cases in India)

A. brunneoviolaceus (=A. fijiensis)

New species of Aspergillus section Nigri

A. floridensis A. trinidadensis

A. brunneoviolaceus A. uvarum (syn. A.fijiensis)

Jurjević Ž. et al. 2012. IMA Fungus, 3(2): 159–173. 72 Aspergillus section Usti

Aspergillus calidoustus sp. nov.

• All except one clinical “A. ustus” isolates belong to this species • Relatively rare human pathogen (28-30 cases reported) • Able to grow at >37°C • Associated with high mortality rates, primarily due to the reduced susceptibilities to azoles

Eukaryotic Cell 2008; 7: 630-638 A. versicolor A. puniceus Tree based on –tubulin A. ustus sequences A. granulosus growth at 37°C (+) A. keveii A. insuetus A. pseudodeflectus growth at 37°C (++)

clinical isolates; A. calidoustus growth at 37°C (++)

Hülle-cells Stud Mycol 2007; 59:107-128 Incidence of species of section Usti in indoor environments

A. pseudodeflectus A. ustus 4% A. keveii 4% 7% A. germanicus 4%

A. puniceus A. calidoustus 22% 59%

Aspergillus section Circumdati

Ochratoxin A DTO 28-E8 A. ochraceus DTO 31-F6 A. ochraceus DTO 48-F3 A. ochraceus DTO 48-F7 A. ochraceus DTO 48-G2 A. ochraceus DTO 11-F2 A. ochraceus A. westerdijkiae CBS 112803 DTO 46-D7 A. ochraceus DTO 50-H4 A. ochraceus DTO 50-H5 A. ochraceus DTO 50-I4 A. ochraceus A. westerdijkiae CBS 112791 DTO 46-D9 A. ochraceus A. westerdijkiae CBS 112804 DTO 17-B7 A. ochraceus DTO 31-F8 A. ochraceus DTO 46-E1 A. ochraceus DTO 26-A1 A. ochraceus DTO 48-F1 A. ochraceus 65 DTO 7-D3 A. ochraceus DTO 4-I6 A. ochraceus DTO 25-H8 A. ochraceus DTO 46-D8 A. ochraceus DTO 8-H3 A. ochraceus DTO 50-H9 A. ochraceus DTO 50-G8 A. ochraceus DTO 25-I3 A. ochraceus DTO 50-G9 A. ochraceus DTO 48-G3 A. ochraceus 99 DTO 48-F4 A. ochraceus DTO 48-F5 A. ochraceus DTO 50-G4 A. ochraceus DTO 50-G6 A. ochraceus DTO 50-G5 A. ochraceus 67 DTO 13-D2 A. ochraceus DTO 50-I3 A. ochraceus DTO 28-E7 A. ochraceus

48 DTO 50-I5 A. ochraceus DTO 48-F8 A. ochraceus DTO 48-G5 A. ochraceus 95 DTO 50-G7 A. ochraceus A. ostianus CBS 54865 98 DTO 50-H3 A. ochraceus A. ostianus CBS 31180 DTO 48-F2 A. ochraceus DTO 50-G3 A. ochraceus

57 A. ochraceus CBS 10808 66 DTO 48-E8 A. ochraceus 95 DTO 48-F6 A. ochraceus A. petrakii/ochraceus CBS 10557 90 DTO 48-E6 A. ochraceus DTO 48-G4 A. ochraceus A. ochraceus CBS 62478 DTO 50-H1 A. ochraceus A. ochraceus CBS 74870 63 DTO 15-E7 A. ochraceus 65 DTO 48-E4 A. ochraceus

100 A. melleus CBS 112786 A. melleus CBS 54665

92 DTO 47-F2 A. ochraceus … 84 … … A. petrakii/melleus CBS 64078 66 DTO 28-E8 A. ochraceus

78 A. melleus CBS 11426

DTO 31-F6 A. ochraceus A. cretensis CBS 112802 DTO 48-F3 A. ochraceus 100 A. cretensis CBS 112805 A. cretensis 57 DTO 48-F7 A. ochraceus Petromyces muricatus CBS 112808 P. muricatus DTO 48-G2 A. ochraceus 100 Petromyces muricatus CBS 112809 29 DTO 11-F2 A. ochraceus 61 Petromyces muricatus CBS 112810 A. westerdijkiae CBS 112803 DTO 31-F5 A. ochraceus Aspergillus sp. (1) DTO 46-D7 A. ochraceus A. auricomus CBS 46765 DTO 50-H4 A. ochraceus 100 A. auricomus CBS 61378 A. auricomus DTO 50-H5 A. ochraceus A. steynii CBS 112814 DTO 50-I4 A. ochraceus 47 DTO 31-F7 A. ochraceus A. westerdijkiae CBS 112791 91 A. steynii CBS 112812 A. steynii (4) DTO 46-D9 A. ochraceus DTO 28-E6 A. ochraceus A. westerdijkiae CBS 112804 100 DTO 47-F1 A. ochraceus DTO 17-B7 A. ochraceus DTO 47-E9 A. ochraceus DTO 31-F8 A. ochraceus A. elegans CBS 10214 A. elegans DTO 46-E1 A. ochraceus 96 A. insulicola CBS 38275 A. insulicola DTO 26-A1 A. ochraceus 98 A. pseudoelegans CBS 112796 DTO 48-F1 A. ochraceus 65 49 A. pseudoelegans CBS 112797 A. pseudoelegans DTO 7-D3 A. ochraceus 100 A. ochraceopetaliformis CBS 12355 DTO 4-I6 A. ochraceus DTO 25-H8 A. ochraceus 47 A. flocculosus CBS 116336 A. westerdijkiae (36) A. flocculosus CBS 112798 DTO 46-D8 A. ochraceus DTO 8-H3 A. ochraceus 98 A. flocculosus CBS 112784 A. flocculosus DTO 50-H9 A. ochraceus A. flocculosus CBS 112799 DTO 50-G8 A. ochraceus A. flocculosus CBS 112789 DTO 25-I3 A. ochraceus 63 A. flocculosus CBS 112785 DTO 50-G9 A. ochraceus A. roseoglobulosus CBS 112800 A. roseoglobulosus DTO 48-G3 A. ochraceus 71 A. neobridgeri CBS 55982 99 DTO 48-F4 A. ochraceus DTO 13-I8 A. ochraceus A. nebridgeri DTO 48-F5 A. ochraceus DTO 48-E7 A. ochraceus DTO 50-G4 A. ochraceus DTO 13-D1 A. ochraceus DTO 50-G6 A. ochraceus 79 DTO 50-H6 A. ochraceus A. persii I (8) DTO 50-G5 A. ochraceus 67 DTO 17-B8 A. ochraceus DTO 13-D2 A. ochraceus DTO 13-C9 A. ochraceus DTO 50-I3 A. ochraceus DTO 28-E9 A. ochraceus DTO 28-E7 A. ochraceus DTO 13-I7 A. ochraceus A. bridgeri 48 DTO 50-I5 A. ochraceus A. bridgeri CBS 35081 DTO 48-F8 A. ochraceus DTO 48-F9 A. ochraceus DTO 48-G5 A. ochraceus 99 A. persii CBS 112795 95 DTO 50-G7 A. ochraceus DTO 50-I1 A. ochraceus A. persii II (5) A. ostianus CBS 54865 98 DTO 50-F9 A. ochraceus DTO 50-H3 A. ochraceus A. ostianus (1) DTO 50-H2 A. ochraceus A. ostianus CBS 31180 DTO 48-G1 A. ochraceus DTO 48-F2 A. ochraceus A. sulphureus CBS 55065 DTO 50-G3 A. ochraceus 61 99 A. sulphureus CBS 38575 A. sulphureus 57 A. ochraceus CBS 10808 66 A. sclerotiorum CBS 63278 DTO 48-E8 A. ochraceus 95 DTO 13-D4 A. ochraceus DTO 48-F6 A. ochraceus A. petrakii/ochraceus CBS 10557 DTO 28-F1 A. ochraceus 90 DTO 48-E6 A. ochraceus DTO 50-H8 A. ochraceus A. ochraceus (9) DTO 48-G4 A. ochraceus A. sclerotiorum CBS 54965 A. ochraceus CBS 62478 DTO 48-E5 A. ochraceus DTO 50-H1 A. ochraceus 97 DTO 50-G2 A. ochraceus A. sclerotiorum (10) A. ochraceus CBS 74870 DTO 13-D3 A. ochraceus 63 DTO 15-E7 A. ochraceus DTO 48-E9 A. ochraceus 65 DTO 48-E4 A. ochraceus DTO 50-I2 A. ochraceus 100 A. melleus CBS 112786 DTO 50-G1 A. ochraceus A. melleus CBS 54665 DTO 50-H7 A. ochraceus 92 DTO 47-F2 A. ochraceus A. melleus (1) DTO 48-G6 A. terreus 84 66 A. petrakii/melleus CBS 64078 78 A. melleus CBS 11426 10 A. cretensis CBS 112802

57 100 A. cretensis CBS 112805 Petromyces muricatus CBS 112808

100 Petromyces muricatus CBS 112809 29 61 Petromyces muricatus CBS 112810 DTO 31-F5 A. ochraceus A. auricomus CBS 46765 100 A. auricomus CBS 61378 A. steynii CBS 112814 47 DTO 31-F7 A. ochraceus

91 A. steynii CBS 112812 DTO 28-E6 A. ochraceus 100 DTO 47-F1 A. ochraceus DTO 47-E9 A. ochraceus A. elegans CBS 10214

96 A. insulicola CBS 38275

98 A. pseudoelegans CBS 112796 49 A. pseudoelegans CBS 112797 100 A. ochraceopetaliformis CBS 12355

47 A. flocculosus CBS 116336 A. flocculosus CBS 112798

98 A. flocculosus CBS 112784 A. flocculosus CBS 112799 A. flocculosus CBS 112789 63 A. flocculosus CBS 112785 A. roseoglobulosus CBS 112800 71 A. neobridgeri CBS 55982 DTO 13-I8 A. ochraceus DTO 48-E7 A. ochraceus DTO 13-D1 A. ochraceus

79 DTO 50-H6 A. ochraceus DTO 17-B8 A. ochraceus DTO 13-C9 A. ochraceus DTO 28-E9 A. ochraceus DTO 13-I7 A. ochraceus A. persii CBS 112795 DTO 48-F9 A. ochraceus 99 A. bridgeri CBS 35081 DTO 50-I1 A. ochraceus DTO 50-F9 A. ochraceus DTO 50-H2 A. ochraceus DTO 48-G1 A. ochraceus A. sulphureus CBS 55065 61 99 A. sulphureus CBS 38575 A. sclerotiorum CBS 63278 DTO 13-D4 A. ochraceus DTO 28-F1 A. ochraceus DTO 50-H8 A. ochraceus A. sclerotiorum CBS 54965 DTO 48-E5 A. ochraceus 97 DTO 50-G2 A. ochraceus DTO 13-D3 A. ochraceus DTO 48-E9 A. ochraceus DTO 50-I2 A. ochraceus DTO 50-G1 A. ochraceus DTO 50-H7 A. ochraceus DTO 48-G6 A. terreus

10 Incidence of species of section Circumdati in indoor environments A. melleus A. ostianus 2% 1% A. steynii A. westlandii A. ochraceus 5% 1% 12%

A. westerdijkiae A. sclerotiorum 48% 14%

A. persii 17% A. ochraceopetaliformis in an onychomycosis case

Med Mycol

Aspergillus persii Aspergillus melleus 2007: IV finger foot dx, direct examination 57 year old, male

Aspergillus melleus A. westerdijkiae and A. insulicola are also able to cause onychomycoses

Med Mycol 50 (2013) 601

Aspergillus section Terrei

lovastatin OH O

O O

O

Lovastatin Aspergillus terreus

• 3rd most frequent cause of IA worldwide; the most frequent causative agent of IA in some medical centers (eg. Austria) • Increased lethality of infections compared with those caused by other Aspergillus species • Construction activity, soil of potted plants, and water distribution systems implicated as potential reservoirs of A. terreus in hospital environments • Exhibit decreased in vitro susceptibilities to amphotericin B

Aspergilli in IA cases in Austria

Distribution of Aspergilli from IA cases at Innsbruck University, Austria (2003-2006)

A. flavus 6% A. fumigatus 36%

A. terreus 58%

Clin Infect Dis 45 (2007) e101-e104 A. alabamensis sp. nov.

• Multilocus sequence analysis of large numbers of (mainly) clinical isolates revealed a new pathogenic species related to A. terreus • Occurrence: USA (Alabama), Argentina • Mostly recovered as colonizing isolates from immunocompetent populations • Exhibit decreased in vitro susceptibilities to amphotericin B

A. alabamensis sp. nov.

Figure 5 F. flavipes NRRL6134Aniveusvarindicus 100 NRRL4101Aallahabadii 98 NRRL4539Aallahabadii 91 98 NRRL5505Aniveus NRRL4751Aniveus NRRL515Aniveus A. terreus var aureus IBT22563 UAB18 C Alabama 100 CBS15F8 E Argentina CBS15F9 E Argentina UAB22 C Alabama 97 UAB33 C Alabama UAB28 C Alabama UAB1 C Alabama UAB20 C Alabama Clade 2 UAB23 C Alabama UAB13 C Alabama 100 UAB15 C Alabama UAB30 C Alabama UAB38 C Alabama CBS134 60 E CBS6I9 E Holland IBT6271 UAB4 C Alabama IBT16744 E Galapagos IBT16745 E Galapagos IBT26384 C Brazil 100 UAB5 C Alabama CBS469 81 C Thailand 1686 05 C India CBS383 75 E India CBS130.55 A. terreus var africanus IBT13121 NRRL4609 A terreus var africanus 1796 05 C India CBS19F3 C India UAB34 C Alabama UAB26 C Alabama CBS2B7 E Holland IBT26385 IBT14590 IBT6450 IBT24859 E Slovenia CBS118 27 P32 C Austria UAB19 C Alabama UAB7 C Alabama P26 C Austria IBT20944 UAB17 C Alabama CBS125 38 E NewZealand CBS594 65 E E8 E Austria Clade 1 E7 E Austria E5 E Austria UAB9 C Alabama UAB31 C Alabama P10 C Austria CBS601 65 E Connecticut P9 C Austria CBS106 25 CBS17A1 E Holland P7 C Austria CBS8G3 E Holland P29 C Austria P30 C Austria IBT15722 E9 E Austria A terreusNIH2624 P11 C Austria P33 C Austria P38 C Austria P23 C Austria CBS8G5 E Holland P20 C Austria IBT12713 O New Mexico IBT13089 IBT21125 P15 C Austria 1769 05 C India IBT26915 O Panama UAB6 C Alabama UAB11 C Alabama UAB10 C Alabama CBS24A3 E Holland CBS24A4 E Holland P16 C Austria IBT23544 E P14 C Austria UAB32 C Alabama UAB2 C Alabama UAB37 C Alabama P22 C Austria UAB21 C Alabama P13 C Austria 0.005 substitutions/site P12 C Austria P4 C Austria UAB8 C Alabama UAB12 C Alabama Eukaryotic Cell (submitted) UAB3 C Alabama Aspergillus hortai

• Originally described in 1935 from ear infection (Brazil) • Also identified from nail infection in the Czech Republic

Med Mycol 50 (2013) 601 Aspergillus section Candidi

Neighbor-joining tree based on -tubulin sequence data of Aspergillus section Candidi

A. flavus CBS 100927 ( EF203132) A. tritici 11-H7 (Netherlands) 100 A. tritici CBS 117270 (drums, Ghana) A. tritici CBS 119225 (flour, Sweden) A. tritici SZMC 0897 (Hungary) A. tritici SZMC 0565 (Hungary) A. triticiT CBS 266.81 (wheat, India) 85 A. thaichungensisT IBT 19404 (Taiwan) A. candidus CBS 567.65 (Brazil) 63/497 PI char. T 70 A. campestris CBS 348.81 (USA) 4 MP trees A. candidus CBS 225.80 (nail, Netherlands) 100 L: 218 100 A. candidus 13-C4 (Netherlands) A. candidus CBS 116945 (dust, Netherlands) CI: 0.8853 A. candidus 25-I1 (Germany) RI: 0.9178 A. candidus 1-F9 (Netherlands) 91 T 100 A. candidus CBS 566.65 (?) A. candidusT IMI 091889 (?) 100 A. candidus 17-C2 (Netherlands) A. candidus CBS 175.68 (mouse dung, Netherlands)

98 A. candidus CBS 114385 (air, Finland) A. candidus CBS 119.28 (Japan) A. candidus CBS 102.13 (Japan) A. candidus CBS 118.28 (?)

0.1 Aspergillus tritici

• Originally described as “A. triticus” by Mehrotra & Basu (1976) • Isolates grow well at 37ºC • More yellowish than A. candidus • Some of them produce purple sclerotia • Conidial heads radiate, wet (“slimy”), conidia roughened • Causes onychomycoses (Hubka et al. Med. Mycol. 50: 601) A. pragensis

• Potential cause of nail infection in humans • Does not grow at 37°C • Isolated in the Czech Republic

Hubka et al. submitted Aspergillus section Nidulantes A. nidulans A. quadrilineata

E. rugulosa E. nidulans var. echinulata Emericella nidulans (Aspergillus nidulans) • Important model organism • Uncommon in animals and humans • Occurs predominantly in patients with chronic granulomatous disease (CGD)

E. heterothallicaT CBS 489.65 E. nidulans var. echinulataT CBS 120.55 100 4606 V44-73 E. nidulans var. echinulata V53-70

95 E. rugulosa CBS 198.88 E. rugulosa CBS 171.71 E. rugulosa E. rugulosa CBS 199.88 V43-77 E. quadrilineataT CBS 591.65 Tree based on V42-23 V42-81 calmodulin sequences 100 100 V44-47 V42-43 V24-62 V43-21 E. quadrilineata V49-18 E. nidulans CBS 291.95 • 10/22 E. quadrilineata V40-61 90 V43-63 • 8/22 E. nidulans V49-25 E. nidulans CBS 121.35 • 3/22 E. nidulans var. echinulata E. nidulansT CBS 589.65 E. nidulans CBS 288.95 100 V45-01 • 1/22 E. rugulosa V32-12 V44-45 V44-46 E. nidulans V44-48 V06-20 9043 E. nidulans CBS 565.70 0.1 9059

Emerg Infect Dis 2008; 14: 566-572 Incidence of Emericella species among the examined clinical isolates

Emerg Infect Dis 2008; 14: 566-572 Comparison of mean MICs against E. nidulans (n=12) and E. quadrilineata (n=12)

Drug E. nidulans E. quadrilineata Significance

Amphotericin B 2.5 0.5 P<0.05

Itraconazole 0.07 0.13 N.S.

Voriconazole 0.26 0.39 P<0.05

Posaconazole 0.25 0.22 P<0.05

Caspofungin 0.32 1.83 P<0.05

Terbinafin 0.01 0.009 N.S.

Emerg Infect Dis 2008; 14: 566-572

A. rugulovalvus (=Emericella rugulosa) Aspergillus section Versicolores A. creber A. cvjetkovici A. fructus A. jensenii

A. puulaauensis A. southafricanus A. subversicolor A. tennesseensis

A. toxicarium A. amoenus A. protuberus A. versicolor

Jurjević Ž. et al. 2012. IMA Fungus, 3(1): 59-79.

A. sydowii on gorgonian sea fans

Collaborators

– Sándor Kocsubé, Gyöngyi Szigeti, Nikolett Baranyi, University of Szeged, Szeged, Hungary – Robert A. Samson, CBS-KNAW Fungal Biodiversity Centre, Utrecht, Netherlands – Jens C. Frisvad, Technical University, Lyngby, Denmark – Vit Hubka, Vanessa Barrs, Zeljko Jurjevic, Mirca Zotti Thanks for your attention!