BackgroundAspergillusspecies are important causes of invasive fungal disease, particularly among those with an impaired immune system. Increasing reports have revealed a rising incidence of antifungal drug resistance amongAspergillusspp., particularly among cryptic species. Understanding local antifungal susceptibility patterns is paramount to delivering optimal clinical care.MethodsAspergillusspp. recovered from clinical specimens between 2000 and 2021 from Pathology Queensland were collected.Aspergillusspp. were identified routinely morphologically, and where there was ambiguity or a lack of sporulation, by sequencing of the internal transcribed spacer (ITS) region. AllAspergillusspp. that underwent antifungal susceptibility testing according to the CLSI M38-A3 method and were recorded and included in the study. Amphotericin B, voriconazole, posaconazole, isavuconazole, micafungin, caspofungin, and anidulafungin were tested. Pathology Queensland services all public healthcare facilities in Queensland, Australia.Results236Aspergillusspp. were identified from clinical specimens during the study period. The most frequent species identified wereAspergillussection Fumigati (n= 119),Aspergillussection Flavi (n= 35),Aspergillus terreus(n= 32) andAspergillus niger(n= 29). Overall, MIC50/90values for voriconazole, posaconazole, itraconazole, and isavuconazole were 0.25/1, 0.25/0.5, 0.25/0.5, and 0.5/2 mg/L respectively. Echinocandins demonstrated low MIC values overall with micafungin and anidulafungin both having an MIC50/90of 0.015/0.03 mg/L. A total of 15 cryptic species were identified; high triazole MIC values were observed with a voriconazole MIC50/90of 2/8 mg/L. From 2017 to 2021 we observed an increase in incidence of isolates with high voriconazole MIC values. There was no difference in voriconazole MIC values betweenAspergillusspp. acquired in North Queensland when compared to Southeast Queensland, Australia.ConclusionIncreasing reports of antifungal resistance amongAspergillusspp. is concerning and warrants further investigation both locally and worldwide. Active surveillance of both the emergence of differentAspergillusspp. and changes in antifungal susceptibility patterns over time is crucial to informing clinicians and treatment guidelines.