title: Cryptic species and azole resistance in the Aspergillus niger complex.
abstract: Aspergillus niger is a common clinical isolate. Multiple species comprise the Aspergillus section Nigri and are separable using sequence data. The antifungal susceptibility of these cryptic species is not known. We determined the azole MICs of 50 black aspergilli, 45 from clinical specimens, using modified EUCAST (mEUCAST) and Etest methods. Phylogenetic trees were prepared using the internal transcribed spacer, beta-tubulin, and calmodulin sequences to identify strains to species level and the results were compared with those obtained with cyp51A sequences. We attempted to correlate cyp51A mutations with azole resistance. Etest MICs were significantly different from mEUCAST MICs (P < 0.001), with geometric means of 0.77 and 2.79 mg/liter, respectively. Twenty-six of 50 (52%) isolates were itraconazole resistant by mEUCAST (MICs > 8 mg/liter), with limited cross-resistance to other azoles. Using combined beta-tubulin/calmodulin sequences, the 45 clinical isolates grouped into 5 clades, A. awamori (55.6%), A. tubingensis (17.8%), A. niger (13.3%), A. acidus (6.7%), and an unknown group (6.7%), none of which were morphologically distinguishable. Itraconazole resistance was found in 36% of the isolates in the A. awamori group, 90% of the A. tubingensis group, 33% of the A. niger group, 100% of the A. acidus group, and 67% of the unknown group. These data suggest that cyp51A mutations in section Nigri may not play as important a role in azole resistance as in A. fumigatus, although some mutations (G427S, K97T) warrant further study. Numerous cryptic species are found in clinical isolates of the Aspergillus section Nigri and are best reported as "A. niger complex" by clinical laboratories. Itraconazole resistance was common in this data set, but azole cross-resistance was unusual. The mechanism of resistance remains obscure.
authors.affiliation: The University of Manchester, 1.800 Stopford Building, Oxford Road, Manchester M13 9PT, United Kingdom. susan.j.howard@manchester.ac.uk
authors.forename: Susan J
authors.initials: SJ
authors.lastname: Howard
authors.forename: Elizabeth
authors.initials: E
authors.lastname: Harrison
authors.forename: Paul
authors.initials: P
authors.lastname: Bowyer
authors.forename: Janos
authors.initials: J
authors.lastname: Varga
authors.forename: David W
authors.initials: DW
authors.lastname: Denning
mesh_terms: D000935:Antifungal Agents / Q000494:pharmacology*; D001228:Aspergillosis / Q000188:drug therapy / Q000382:microbiology; D001234:Aspergillus niger / Q000187:drug effects* / Q000235:genetics / Q000302:isolation & purification; D001393:Azoles / Q000494:pharmacology*; D001483:Base Sequence; D002147:Calmodulin / Q000235:genetics; D003577:Cytochrome P-450 Enzyme System / Q000235:genetics; D021901:DNA, Intergenic / Q000235:genetics; D025141:Drug Resistance, Fungal / Q000235:genetics*; D005656:Fungal Proteins / Q000235:genetics; D017964:Itraconazole / Q000494:pharmacology; D008826:Microbial Sensitivity Tests; D009154:Mutation; D010802:Phylogeny; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D014404:Tubulin / Q000235:genetics
chemical_list: D000935:Antifungal Agents; D001393:Azoles; D002147:Calmodulin; D021901:DNA, Intergenic; D005656:Fungal Proteins; D014404:Tubulin; D017964:Itraconazole; D003577:Cytochrome P-450 Enzyme System; C465526:cytochrome P-450 CYP51A, Aspergillus