Effects of Agricultural Fungicide Use on Aspergillus fumigatus Abundance, Antifungal Susceptibility, and Population Structure

Abstract

Antibiotic resistance is an increasing threat to human health. In the case of Aspergillus fumigatus, which is both an environmental saprobe and an opportunistic human fungal pathogen, resistance is suggested to arise from fungicide use in agriculture, as the azoles used for plant protection share the same molecular target as the frontline antifungals used clinically. However, limiting azole fungicide use on crop fields to preserve their activity for clinical use could threaten the global food supply via a reduction in yield. In this study, we clarify the link between azole fungicide use on crop fields and resistance in a prototypical human pathogen through systematic soil sampling on farms in Germany and surveying fields before and after fungicide application. We observed a reduction in the abundance of A. fumigatus on fields following fungicide treatment in 2017, a finding that was not observed on an organic control field with only natural plant protection agents applied. However, this finding was less pronounced during our 2018 sampling, indicating that the impact of fungicides on A. fumigatus population size is variable and influenced by additional factors. The overall resistance frequency among agricultural isolates is low, with only 1 to 3% of isolates from 2016 to 2018 displaying resistance to medical azoles. Isolates collected after the growing season and azole exposure show a subtle but consistent decrease in susceptibility to medical and agricultural azoles. Whole-genome sequencing indicates that, despite the alterations in antifungal susceptibility, fungicide application does not significantly affect the population structure and genetic diversity of A. fumigatus in fields. Given the low observed resistance rate among agricultural isolates as well the lack of genomic impact following azole application, we do not find evidence that azole use on crops is significantly driving resistance in A. fumigatus in this context. IMPORTANCE Antibiotic resistance is an increasing threat to human health. In the case of Aspergillus fumigatus, which is an environmental fungus that also causes life-threatening infections in humans, antimicrobial resistance is suggested to arise from fungicide use in agriculture, as the chemicals used for plant protection are almost identical to the antifungals used clinically. However, removing azole fungicides from crop fields threatens the global food supply via a reduction in yield. In this study, we survey crop fields before and after fungicide application. We find a low overall azole resistance rate among agricultural isolates, as well as a lack of genomic and population impact following fungicide application, leading us to conclude azole use on crops does not significantly contribute to resistance in A. fumigatus.

Dynamics of Aspergillus fumigatus in Azole Fungicide-Containing Plant Waste in the Netherlands (2016–2017)

“…This is in contrast to studies where samples were taken at a location without azole pressure, where 10 3 CFU/g of resistant A. fumigatus was found ( 8 ). In German organic fields without fungicides exposure, 0 to 1 CFU/g displayed resistance to medical azoles ( 14 ). Thus, in our study, we provide data backing up previous suggestions that azole-containing decaying plant material is a reservoir and selective environment for resistant A. fumigatus .…”

Sterol 14α-Demethylase Ligand-Binding Pocket-Mediated Acquired and Intrinsic Azole Resistance in Fungal Pathogens

“…Triazole antifungals were first licensed for use in humans in the 1990s. Today, five triazole drugs are available for therapy of systemic fungal infections: fluconazole (FLC), itraconazole (ITC), voriconazole (VCZ), posaconazole (PCZ), and isavuconazole (IVU) [ 4 , 5 ]. The azoles are used extensively because they are significantly less expensive than other antifungal treatments and their overall antimycotic activity is relatively broad spectrum [ 6 ].…”

“…With fungal pathogens estimated to cause yield losses of up to 30%, the use of fungicides is essential for efficient food production and overall food security [ 5 ]. Because azole drugs are amongst the best-tolerated and most active antifungals, they are often the first line treatment in human and veterinary medicine for many fungal diseases [ 4 , 16 ].…”

“…Their popularity and extensive use comes at a cost due to the resultant positive selection pressure for azole-resistance amongst fungal pathogens of plants, animals, and humans [ 17 ]. An increased incidence of intrinsically azole-resistant fungi and the emergence azole-resistant strains from naturally susceptible fungi has resulted in major losses in food production and in therapeutic failures in both veterinary and human medicine [ 5 ]. Many pathogenic molds of humans are saprophytes that decay organic matter and are ubiquitous in the environment, including agricultural soils.…”

Hospital Environment as a Source of Azole-Resistant Aspergillus fumigatus Strains with TR34/L98H and G448S Cyp51A Mutations

“…Azole-resistant A. fumigatus isolates from clinical and environmental origin are being acknowledged worldwide [ 10 , 14 ]. Resistant strainsharbordifferent resistance mechanisms that confer particular susceptibility profiles to antifungal drugs used in the clinical setting and also in the environment [ 16 , 43 ]. To date, the most prevalent azole resistance mechanism is the TR34/L98H combination in Cyp51A, the target for azole drugs, which has been mostly described from environmental isolates conferring multi-azole resistance [ 10 ].…”