Cryptic species of Aspergillus fumigatus, including the Aspergillus viridinutans species complex, are increasingly reported to be causes of invasive aspergillosis. Their identification is clinically relevant, as these species frequently have intrinsic resistance to common antifungals. We evaluated the susceptibilities of 90 environmental and clinical isolates from the A. viridinutans species complex, identified by DNA sequencing of the calmodulin gene, to seven antifungals (voriconazole, posaconazole, itraconazole, amphotericin B, anidulafungin, micafungin, and caspofungin) using the reference European Committee on Antimicrobial Susceptibility Testing (EUCAST) method. The majority of species demonstrated elevated MICs of voriconazole (geometric mean [GM] MIC, 4.46 mg/liter) and itraconazole (GM MIC, 9.85 mg/liter) and had variable susceptibility to amphotericin B (GM MIC, 2.5 mg/liter). Overall, the MICs of posaconazole and the minimum effective concentrations of echinocandins were low. The results obtained by the EUCAST method were compared with the results obtained with Sensititre YeastOne (YO) panels. Overall, there was 67% agreement (95% confidence interval [CI], 62 to 72%) between the results obtained by the EUCAST method and those obtained with YO panels when the results were read at 48 h and 82% agreement (95% CI, 78 to 86%) when the results were read at 72 h. There was a significant difference in agreement between antifungals; agreement was high for amphotericin B, voriconazole, and posaconazole (70 to 86% at 48 h and 88 to 93% at 72 h) but was very low for itraconazole (37% at 48 h and 57% at 72 h). The agreement was also variable between species, with the maximum agreement being observed for A. felis isolates (85 and 93% at 48 and 72 h, respectively). Elevated MICs of voriconazole and itraconazole were cross-correlated, but there was no correlation between the other azoles tested.

Department of Botany Faculty of Science Charles University Prague Czech Republic

Department of Microbiology Faculty of Medicine and Dentistry Palacky University Olomouc Czech Republic

Department of Nutrition Science University of Nagasaki Nagasaki Japan

Laboratory of Clinical Mycology Institute of Public Health Ostrava Czech Republic

Laboratory of Fungal Genetics and Metabolism Institute of Microbiology of the CAS Prague Czech Republic

Laboratory of Medical Mycology Department of Parasitology Mycology and Mycobacteriology Prague Public Health Institute in Usti nad Labem Prague Czech Republic

Medical Mycology Research Center Chiba University Chiba Japan

Sydney School of Veterinary Science and Marie Bashir Institute of Infectious Diseases and Biosecurity University of Sydney Camperdown New South Wales Australia

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Lin SJ, Schranz J, Teutsch SM. 2001. Aspergillosis case-fatality rate: systematic review of the literature. Clin Infect Dis 32:358–366. doi: 10.1086/318483. PubMed DOI

Alastruey-Izquierdo A, Mellado E, Peláez T, Pemán J, Zapico S, Alvarez M, Rodríguez-Tudela JL, Cuenca-Estrella M, FILPOP Study Group. 2013. Population-based survey of filamentous fungi and antifungal resistance in Spain (FILPOP study). Antimicrob Agents Chemother 57:3380–3387. doi: 10.1128/AAC.00383-13. PubMed DOI PMC

Lamoth F. 2016. PubMed DOI PMC

Balajee SA, Gribskov JL, Hanley E, Nickle D, Marr KA. 2005. PubMed DOI PMC

Sugui JA, Vinh DC, Nardone G, Shea YR, Chang YC, Zelazny AM, Marr KA, Holland SM, Kwon-Chung KJ. 2010. PubMed DOI PMC

Nováková A, Hubka V, Dudová Z, Matsuzawa T, Kubátová A, Yaguchi T, Kolařík M. 2014. New species in DOI

Yaguchi T, Horie Y, Tanaka R, Matsuzawa T, Ito J, Nishimura K. 2007. Molecular phylogenetics of multiple genes on PubMed DOI

Balajee SA, Kano R, Baddley JW, Moser SA, Marr KA, Alexander BD, Andes D, Kontoyiannis DP, Perrone G, Peterson S, Brandt ME, Pappas PG, Chiller T. 2009. Molecular identification of PubMed DOI PMC

Barrs VR, van Doorn TM, Houbraken J, Kidd SE, Martin P, Pinheiro MD, Richardson M, Varga J, Samson RA. 2013. PubMed DOI PMC

Eamvijarn A, Manoch L, Chamswarng C, Piasai O, Visarathanonth N, Luangsaard JJ, Kijjoa A. 2013. DOI

Barrs V, Beatty J, Dhand NK, Talbot J, Bell E, Abraham L, Chapman P, Bennett S, van Doorn T, Makara M. 2014. Computed tomographic features of feline sino-nasal and sino-orbital aspergillosis. Vet J 201:215–222. doi: 10.1016/j.tvjl.2014.02.020. PubMed DOI

Sugui JA, Peterson SW, Figat A, Hansen B, Samson RA, Mellado E, Cuenca-Estrella M, Kwon-Chung KJ. 2014. Genetic relatedness versus biological compatibility between PubMed DOI PMC

Matsuzawa T, Takaki GMC, Yaguchi T, Okada K, Abliz P, Gonoi T, Horie Y. 2015. DOI

Shigeyasu C, Yamada M, Nakamura N, Mizuno Y, Sato T, Yaguchi T. 2012. Keratomycosis caused by PubMed DOI

Katz ME, Dougall AM, Weeks K, Cheetham BF. 2005. Multiple genetically distinct groups revealed among clinical isolates identified as atypical PubMed DOI PMC

Alcazar-Fuoli L, Mellado E, Alastruey-Izquierdo A, Cuenca-Estrella M, Rodriguez-Tudela JL. 2008. PubMed DOI PMC

Vinh DC, Shea YR, Jones PA, Freeman AF, Zelazny A, Holland SM. 2009. Chronic invasive aspergillosis caused by PubMed DOI PMC

Vinh DC, Shea YR, Sugui JA, Parrilla-Castellar ER, Freeman AF, Campbell JW, Pittaluga S, Jones PA, Zelazny A, Kleiner D, Kwon-Chung KJ, Holland SM. 2009. Invasive aspergillosis due to PubMed DOI PMC

Kano R, Itamoto K, Okuda M, Inokuma H, Hasegawa A, Balajee SA. 2008. Isolation of PubMed DOI

Coelho D, Silva S, Vale-Silva L, Gomes H, Pinto E, Sarmento A, Pinheiro MD. 2011. PubMed DOI

Posteraro B, Mattei R, Trivella F, Maffei A, Torre A, De Carolis E, Posteraro P, Fadda G, Sanguinetti M. 2011. Uncommon PubMed DOI PMC

Barrs VR, Halliday C, Martin P, Wilson B, Krockenberger M, Gunew M, Bennett S, Koehlmeyer E, Thompson A, Fliegner R, Hocking A, Sleiman S, O'Brien C, Beatty JA. 2012. Sinonasal and sino-orbital aspergillosis in 23 cats: aetiology, clinicopathological features and treatment outcomes. Vet J 191:58–64. doi: 10.1016/j.tvjl.2011.02.009. PubMed DOI

Gyotoku H, Izumikawa K, Ikeda H, Takazono T, Morinaga Y, Nakamura S, Imamura Y, Nishino T, Miyazaki T, Kakeya H, Yamamoto Y, Yanagihara K, Yasuoka A, Yaguchi T, Ohno H, Miyzaki Y, Kamei K, Kanda T, Kohno S. 2012. A case of bronchial aspergillosis caused by PubMed DOI

Escribano P, Peláez T, Muñoz P, Bouza E, Guinea J. 2013. Is azole resistance in PubMed DOI PMC

Kano R, Shibahashi A, Fujino Y, Sakai H, Mori T, Tsujimoto H, Yanai T, Hasegawa A. 2013. Two cases of feline orbital aspergillosis due to PubMed DOI

Pelaez T, Alvarez-Perez S, Mellado E, Serrano D, Valerio M, Blanco JL, Garcia ME, Munoz P, Cuenca-Estrella M, Bouza E. 2013. Invasive aspergillosis caused by cryptic PubMed DOI

Alastruey-Izquierdo A, Alcazar-Fuoli L, Cuenca-Estrella M. 2014. Antifungal susceptibility profile of cryptic species of PubMed DOI

Álvarez-Pérez S, Mellado E, Serrano D, Blanco JL, Garcia ME, Kwon M, Munoz P, Cuenca-Estrella M, Bouza E, Peláez T. 2014. Polyphasic characterization of fungal isolates from a published case of invasive aspergillosis reveals misidentification of PubMed DOI

Tamiya H, Ochiai E, Kikuchi K, Yahiro M, Toyotome T, Watanabe A, Yaguchi T, Kamei K. 2015. Secondary metabolite profiles and antifungal drug susceptibility of PubMed DOI

Chong GM, Vonk AG, Meis JF, Dingemans GJH, Houbraken J, Hagen F, Gaajetaan GR, van Tegelen DWE, Simons GFM, Rijnders BJA. 2017. Interspecies discrimination of PubMed DOI

Seki A, Yoshida A, Matsuda Y, Kawata M, Nishimura T, Tanaka J, Misawa Y, Nakano Y, Asami R, Chida K, Kikuchi K, Arai T. 2017. Fatal fungal endocarditis by PubMed DOI

Howard SJ. 2014. Multi-resistant aspergillosis due to cryptic species. Mycopathologia 178:435–439. doi: 10.1007/s11046-014-9774-0. PubMed DOI

Diekema DJ, Messer SA, Hollis RJ, Jones RN, Pfaller MA. 2003. Activities of caspofungin, itraconazole, posaconazole, ravuconazole, voriconazole, and amphotericin B against 448 recent clinical isolates of filamentous fungi. J Clin Microbiol 41:3623–3626. doi: 10.1128/JCM.41.8.3623-3626.2003. PubMed DOI PMC

Sabatelli F, Patel R, Mann PA, Mendrick CA, Norris CC, Hare R, Loebenberg D, Black TA, McNicholas PM. 2006. PubMed DOI PMC

van der Linden JW, Arendrup MC, Warris A, Lagrou K, Pelloux H, Hauser PM, Chryssanthou E, Mellado E, Kidd SE, Tortorano AM, Dannaoui E, Gaustad P, Baddley JW, Uekötter A, Lass-Flörl C, Klimko N, Moore CB, Denning DW, Pasqualotto AC, Kibbler C, Arikan-Akdagli S, Andes D, Meletiadis J, Naumiuk L, Nucci M, Melchers WJ, Verweij PE. 2015. Prospective multicenter international surveillance of azole resistance in PubMed DOI PMC

Gautier M, Normand AC, Ranque S. 2016. Previously unknown species of PubMed DOI

Verweij PE, Zhang J, Debets AJ, Meis JF, van de Veerdonk FL, Schoustra SE, Zwaan BJ, Melchers WJ. 2016. In-host adaptation and acquired triazole resistance in PubMed DOI

Verweij PE, Ananda-Rajah M, Andes D, Arendrup MC, Brüggemann RJ, Chowdhary A, Cornely OA, Denning DW, Groll AH, Izumikawa K, Kullberg BJ, Lagrou K, Maertens J, Meis JF, Newton P, Page I, Seyedmousavi S, Sheppard DC, Viscoli C, Warris A, Donnelly JP. 2015. International expert opinion on the management of infection caused by azole-resistant PubMed DOI

Nedel WL, Pasqualotto AC. 2014. Treatment of infections by cryptic PubMed DOI

Hubka V, Nováková A, Kolařík M, Jurjević Ž, Peterson SW. 2015. Revision of PubMed DOI

Peterson SW. 2008. Phylogenetic analysis of PubMed

Hubka V, Lyskova P, Frisvad JC, Peterson SW, Skorepova M, Kolarik M. 2014. PubMed DOI

Réblová M, Hubka V, Thureborn O, Lundberg J, Sallstedt T, Wedin M, Ivarsson M. 2016. From the tunnels into the treetops: new lineages of black yeasts from biofilm in the Stockholm metro system and their relatives among ant-associated fungi in the PubMed DOI PMC

Katoh K, Standley DM. 2013. MAFFT multiple sequence alignment software version 7: improvements in performance and usability. Mol Biol Evol 30:772–780. doi: 10.1093/molbev/mst010. PubMed DOI PMC

Lanfear R, Calcott B, Ho SY, Guindon S. 2012. PartitionFinder: combined selection of partitioning schemes and substitution models for phylogenetic analyses. Mol Biol Evol 29:1695–1701. doi: 10.1093/molbev/mss020. PubMed DOI

Nguyen L-T, Schmidt HA, von Haeseler A, Minh BQ. 2015. IQ-TREE: a fast and effective stochastic algorithm for estimating maximum-likelihood phylogenies. Mol Biol Evol 32:268–274. doi: 10.1093/molbev/msu300. PubMed DOI PMC

Ronquist F, Teslenko M, van der Mark P, Ayres DL, Darling A, Höhna S, Larget B, Liu L, Suchard MA, Huelsenbeck JP. 2012. MrBayes 3.2: efficient Bayesian phylogenetic inference and model choice across a large model space. Syst Biol 61:539–542. doi: 10.1093/sysbio/sys029. PubMed DOI PMC

Arendrup MC, Guinea J, Cuenca-Estrella M, Meletiadis J, Mouton JW, Lagrou K, Howard SJ, and the Subcommittee on Antifungal Susceptibility Testing (AFST) of the ESCMID European Committee for Antimicrobial Susceptibility Testing (EUCAST). 2015. EUCAST method for the determination of broth dilution minimum inhibitory concentrations of antifungal agents for conidia forming moulds version 9.3. EUCAST, Växjö, Sweden, https://www.aspergillus.org.uk/sites/default/files/pictures/Lab_protocols/EUCAST_E_Def_9_3_Mould_testing_definitive_0.pdf.

Hubka V, Barrs V, Dudová Z, Sklenář F, Kubátová A, Matsuzawa T, Yaguchi T, Horie Y, Nováková A, Frisvad JC, Talbot JJ, Kolařík M. Unravelling species boundaries in the PubMed PMC

Current and Future Pathways in Aspergillus Diagnosis

Unravelling species boundaries in the Aspergillus viridinutans complex (section Fumigati): opportunistic human and animal pathogens capable of interspecific hybridization