scholarly journals The Multi-Fungicide Resistance Status of Aspergillus fumigatus Populations in Arable Soils and the Wider European Environment

2020 ◽  
Vol 11 ◽  
Author(s):  
Bart Fraaije ◽  
Sarah Atkins ◽  
Steve Hanley ◽  
Andy Macdonald ◽  
John Lucas
Keyword(s):  
Aspergillus Fumigatus ◽  
Crop Production ◽  
Agricultural Soils ◽  
Acquired Resistance ◽  
Surface Protein ◽  
Health Concern ◽  
Azole Resistance ◽  
Arable Soils ◽  
Resistance Development ◽  
Resistance Evolution

The evolution and spread of pan-azole resistance alleles in clinical and environmental isolates of Aspergillus fumigatus is a global human health concern. The identification of hotspots for azole resistance development in the wider environment can inform optimal measures to counteract further spread by minimizing exposure to azole fungicides and reducing inoculum build-up and pathogen dispersal. We investigated the fungicide sensitivity status of soil populations sampled from arable crops and the wider environment and compared these with urban airborne populations. Low levels of azole resistance were observed for isolates carrying the CYP51A variant F46Y/M172V/E427K, all belonging to a cluster of related cell surface protein (CSP) types which included t07, t08, t13, t15, t19, and t02B, a new allele. High levels of resistance were found in soil isolates carrying CYP51A variants TR34/L98H and TR46/Y121F/T289A, all belonging to CSP types t01, t02, t04B, or t11. TR46/Y121F/M172V/T289A/G448S (CSP t01) and TR46/Y121F/T289A/S363P/I364V/G448S (CSP t01), a new haplotype associated with high levels of resistance, were isolated from Dutch urban air samples, indicating azole resistance evolution is ongoing. Based on low numbers of pan-azole resistant isolates and lack of new genotypes in soils of fungicide-treated commercial and experimental wheat crops, we consider arable crop production as a coldspot for azole resistance development, in contrast to previously reported flower bulb waste heaps. This study also shows that, in addition to azole resistance, several lineages of A. fumigatus carrying TR-based CYP51A variants have also developed acquired resistance to methyl benzimidazole carbamate, quinone outside inhibitor and succinate dehydrogenase (Sdh) inhibitor fungicides through target-site alterations in the corresponding fungicide target proteins; beta-tubulin (F200Y), cytochrome b (G143A), and Sdh subunit B (H270Y and H270R), respectively. Molecular typing showed that several multi-fungicide resistant strains found in agricultural soils in this study were clonal as identical isolates have been found earlier in the environment and/or in patients. Further research on the spread of different fungicide-resistant alleles from the wider environment to patients and vice versa can inform optimal practices to tackle the further spread of antifungal resistance in A. fumigatus populations and to safeguard the efficacy of azoles for future treatment of invasive aspergillosis.

scholarly journals Relevance of heterokaryosis for adaptation and azole-resistance development in Aspergillus fumigatus

2019 ◽  
Vol 286 (1896) ◽  
pp. 20182886 ◽  
Author(s):  
Jianhua Zhang ◽  
Eveline E. Snelders ◽  
Bas J. Zwaan ◽  
Sijmen E. Schoustra ◽  
Ed J. Kuijper ◽  
...  
Keyword(s):  
Aspergillus Fumigatus ◽  
Human Lung ◽  
Spontaneous Mutation ◽  
Acquired Resistance ◽  
Fungal Mycelium ◽  
Azole Resistance ◽  
Resistance Level ◽  
Resistance Development ◽  
Genetic Changes ◽  
Lung Colonization

Aspergillus fumigatus causes a range of diseases in humans, some of which are characterized by fungal persistence. Aspergillus fumigatus , being a generalist saprotroph, may initially establish lung colonization due to its physiological versatility and subsequently adapt through genetic changes to the human lung environment and antifungal treatments. Human lung-adapted genotypes can arise by spontaneous mutation and/or recombination and subsequent selection of the fittest genotypes. Sexual and asexual spores are considered crucial contributors to the genetic diversity and adaptive potential of aspergilli by recombination and mutation supply, respectively. However, in certain Aspergillus diseases, such as cystic fibrosis and chronic pulmonary aspergillosis, A. fumigatus may not sporulate but persist as a network of fungal mycelium. During azole therapy, such mycelia may develop patient-acquired resistance and become heterokaryotic by mutations in one of the nuclei. We investigated the relevance of heterokaryosis for azole-resistance development in A. fumigatus. We found evidence for heterokaryosis of A. fumigatus in patients with chronic Aspergillus diseases. Mycelium from patient-tissue biopsies segregated different homokaryons, from which heterokaryons could be reconstructed. Whereas all variant homokaryons recovered from the same patient were capable of forming a heterokaryon, those from different patients were heterokaryon-incompatible. We furthermore compared heterokaryons and heterozygous diploids constructed from environmental isolates with different levels of azole resistance. When exposed to azole, the heterokaryons revealed remarkable shifts in their nuclear ratio, and the resistance level of heterokaryons exceeded that of the corresponding heterozygous diploids.

scholarly journals Development and Validation of a High-Resolution Melting Assay To Detect Azole Resistance in Aspergillus fumigatus

2017 ◽  
Vol 61 (12) ◽  
Author(s):  
L. Bernal-Martínez ◽  
H. Gil ◽  
O. Rivero-Menéndez ◽  
S. Gago ◽  
M. Cuenca-Estrella ◽  
...  
Keyword(s):  
High Resolution ◽  
Aspergillus Fumigatus ◽  
Tandem Repeats ◽  
High Resolution Melting ◽  
Screening Method ◽  
Melting Curve ◽  
Health Concern ◽  
Azole Resistance ◽  
Content Type ◽  
Selection Of

ABSTRACT The global emergence of azole-resistant Aspergillus fumigatus strains is a growing public health concern. Different patterns of azole resistance are linked to mutations in cyp51A. Therefore, accurate characterization of the mechanisms underlying azole resistance is critical to guide selection of the most appropriate antifungal agent for patients with aspergillosis. This study describes a new sequencing-free molecular screening tool for early detection of the most frequent mutations known to be associated with azole resistance in A. fumigatus. PCRs targeting cyp51A mutations at positions G54, Y121, G448, and M220 and targeting different tandem repeats (TRs) in the promoter region were designed. All PCRs were performed simultaneously, using the same cycling conditions. Amplicons were then distinguished using a high-resolution melting assay. For standardization, 30 well-characterized azole-resistant A. fumigatus strains were used, yielding melting curve clusters for different resistance mechanisms for each target and allowing detection of the most frequent azole resistance mutations, i.e., G54E, G54V, G54R, G54W, Y121F, M220V, M220I, M220T, M220K, and G448S, and the tandem repeats TR34, TR46, and TR53. Validation of the method was performed using a blind panel of 80 A. fumigatus azole-susceptible or azole-resistant strains. All strains included in the blind panel were properly classified as susceptible or resistant with the developed method. The implementation of this screening method can reduce the time needed for the detection of azole-resistant A. fumigatus isolates and therefore facilitate selection of the best antifungal therapy in patients with aspergillosis.

scholarly journals The Effect of Posaconazole, Itraconazole and Voriconazole in the Culture Medium on Aspergillus fumigatus Triazole Resistance

2020 ◽  
Vol 8 (2) ◽  
pp. 285 ◽  
Author(s):  
Martyna Mroczyńska ◽  
Ewelina Kurzyk ◽  
Magdalena Śliwka-Kaszyńska ◽  
Urszula Nawrot ◽  
Marta Adamik ◽  
...  
Keyword(s):  
Gene Expression ◽  
Aspergillus Fumigatus ◽  
Culture Medium ◽  
Acquired Resistance ◽  
Azole Resistance ◽  
Broth Microdilution ◽  
Microdilution Method ◽  
Broth Microdilution Method ◽  
Selection Of

Triazoles are the only compounds used as antibiotics in both medicine and agriculture. The presence of triazoles in the environment can contribute to the acquisition of azole resistance among isolates of Aspergillus fumigatus. The objective of this study was to investigate the effect of A. fumigatus exposure to triazoles on susceptibility to these compounds. Seventeen triazole-resistant and 21 triazole-sensitive A. fumigatus isolates were examined. The isolates were transferred 20 times on the Sabouraud medium supplemented with posaconazole, itraconazole or voriconazole, followed by five times on the medium not supplemented. The minimum inhibitory concentrations of antimycotics were examined according to the EUCAST broth microdilution method after the 20th transfer and also the 25th transfer. In addition, the expression levels of genes mdr1, mdr2, mdr3, atrF, cyp51A and cyp51B were determined. Cultivation of A. fumigatus on media supplemented with posaconazole, itraconazole and voriconazole resulted in the acquisition of resistance to the tested triazoles of all examined isolates. After recultivation on Sabouraud without azoles, most of the isolates lost their acquired resistance. The long-term use of triazole compounds in agriculture may result in the occurrence of triazole resistant A. fumigatus isolates in the environment, not only by induction or selection of mutations in the cyp51A gene, but also by contribution to changes in the gene expression.

scholarly journals Nonrandom Distribution of Azole Resistance across the Global Population of Aspergillus fumigatus

mBio ◽  
2019 ◽  
Vol 10 (3) ◽  
Author(s):  
Thomas R. Sewell ◽  
Jianing Zhu ◽  
Johanna Rhodes ◽  
Ferry Hagen ◽  
Jacques F. Meis ◽  
...  
Keyword(s):  
Genetic Diversity ◽  
Drug Resistance ◽  
Aspergillus Fumigatus ◽  
Pathogenic Fungus ◽  
Crop Protection ◽  
Resistance Mechanisms ◽  
Health Concern ◽  
Azole Resistance ◽  
Content Type ◽  
Global Population

ABSTRACT The emergence of azole resistance in the pathogenic fungus Aspergillus fumigatus has continued to increase, with the dominant resistance mechanisms, consisting of a 34-nucleotide tandem repeat (TR34)/L98H and TR46/Y121F/T289A, now showing a structured global distribution. Using hierarchical clustering and multivariate analysis of 4,049 A. fumigatus isolates collected worldwide and genotyped at nine microsatellite loci using analysis of short tandem repeats of A. fumigatus (STRAf), we show that A. fumigatus can be subdivided into two broad clades and that cyp51A alleles TR34/L98H and TR46/Y121F/T289A are unevenly distributed across these two populations. Diversity indices show that azole-resistant isolates are genetically depauperate compared to their wild-type counterparts, compatible with selective sweeps accompanying the selection of beneficial mutations. Strikingly, we found that azole-resistant clones with identical microsatellite profiles were globally distributed and sourced from both clinical and environmental locations, confirming that azole resistance is an international public health concern. Our work provides a framework for the analysis of A. fumigatus isolates based on their microsatellite profile, which we have incorporated into a freely available, user-friendly R Shiny application (AfumID) that provides clinicians and researchers with a method for the fast, automated characterization of A. fumigatus genetic relatedness. Our study highlights the effect that azole drug resistance is having on the genetic diversity of A. fumigatus and emphasizes its global importance upon this medically important pathogenic fungus. IMPORTANCE Azole drug resistance in the human-pathogenic fungus Aspergillus fumigatus continues to emerge, potentially leading to untreatable aspergillosis in immunosuppressed hosts. Two dominant, environmentally associated resistance mechanisms, which are thought to have evolved through selection by the agricultural application of azole fungicides, are now distributed globally. Understanding the effect that azole resistance is having on the genetic diversity and global population of A. fumigatus will help mitigate drug-resistant aspergillosis and maintain the azole class of fungicides for future use in both medicine and crop protection.

scholarly journals Azole-Resistance Development; How the Aspergillus fumigatus Lifecycle Defines the Potential for Adaptation

2021 ◽  
Vol 7 (8) ◽  
pp. 599
Author(s):  
Jianhua Zhang ◽  
Alfons J. M. Debets ◽  
Paul E. Verweij ◽  
Eveline Snelders
Keyword(s):  
Genetic Variation ◽  
Aspergillus Fumigatus ◽  
Scientific Work ◽  
Azole Resistance ◽  
Resistance Development ◽  
Genetic Changes ◽  
Genomic Changes ◽  
Metabolic Plasticity ◽  
Resistance Selection ◽  
Made In

In order to successfully infect or colonize human hosts or survive changing environments, Aspergillus fumigatus needs to adapt through genetic changes or phenotypic plasticity. The genomic changes are based on the capacity of the fungus to produce genetic variation, followed by selection of the genotypes that are most fit to the new environment. Much scientific work has focused on the metabolic plasticity, biofilm formation or the particular genetic changes themselves leading to adaptation, such as antifungal resistance in the host. Recent scientific work has shown advances made in understanding the natural relevance of parasex and how both the asexual and sexual reproduction can lead to tandem repeat elongation in the target gene of the azoles: the cyp51A gene. In this review, we will explain how the fungus can generate genetic variation that can lead to adaptation. We will discuss recent advances that have been made in the understanding of the lifecycle of A. fumigatus to explain the differences observed in speed and type of mutations that are generated under different environments and how this can facilitate adaptation, such as azole-resistance selection.

scholarly journals First Description of Azole-Resistant Aspergillus fumigatus Due to TR46/Y121F/T289A Mutation in France

2015 ◽  
Vol 59 (7) ◽  
pp. 4331-4335 ◽  
Author(s):  
Rose-Anne Lavergne ◽  
Florent Morio ◽  
Loïc Favennec ◽  
Stéphane Dominique ◽  
Jacques F. Meis ◽  
...  
Keyword(s):  
Public Health ◽  
Cystic Fibrosis ◽  
Aspergillus Fumigatus ◽  
Environmental Samples ◽  
Health Concern ◽  
Azole Resistance ◽  
Public Health Concern ◽  
Content Type ◽  
High Level ◽  
Level Resistance

ABSTRACTAzole resistance inAspergillus fumigatusis an emerging public health concern. Recently, a novel fungicide-driven mutation in thecyp51Agene and its promoter, TR46/Y121F/T289A, leading to high-level resistance to voriconazole has been identified in The Netherlands, Belgium, Germany, Denmark, Tanzania, and India in both clinical and environmental samples. Here we report the first description ofA. fumigatuscarrying this mutation in France, in a cystic fibrosis patient, underlining the need for extensive monitoring ofAspergillusresistance.

scholarly journals Describing Phosphorus Sorption Processes on Volcanic Soil in the Presence of Copper or Silver Engineered Nanoparticles

Minerals ◽  
2021 ◽  
Vol 11 (4) ◽  
pp. 373
Author(s):  
Jonathan Suazo-Hernández ◽  
Erwin Klumpp ◽  
Nicolás Arancibia-Miranda ◽  
Patricia Poblete-Grant ◽  
Alejandra Jara ◽  
...  
Keyword(s):  
Organic Matter ◽  
Crop Production ◽  
Agricultural Soils ◽  
Ph Value ◽  
Consumer Products ◽  
Soil Samples ◽  
Engineered Nanoparticles ◽  
P Availability ◽  
Agricultural Crop ◽  
Phosphorus Sorption

Engineered nanoparticles (ENPs) present in consumer products are being released into the agricultural systems. There is little information about the direct effect of ENPs on phosphorus (P) availability, which is an essential nutrient for crop growthnaturally occurring in agricultural soils. The present study examined the effect of 1, 3, and 5% doses of Cu0 or Ag0 ENPs stabilized with L-ascorbic acid (suspension pH 2–3) on P ad- and desorption in an agricultural Andisol with total organic matter (T-OM) and with partial removal of organic matter (R-OM) by performing batch experiments. Our results showed that the adsorption kinetics data of H2PO4− on T-OM and R-OM soil samples with and without ENPs were adequately described by the pseudo-second-order (PSO) and Elovich models. The adsorption isotherm data of H2PO4− from T-OM and R-OM soil samples following ENPs addition were better fitted by the Langmuir model than the Freundlich model. When the Cu0 or Ag0 ENPs doses were increased, the pH value decreased and H2PO4− adsorption increased on T-OM and R-OM. The H2PO4− desorption (%) was lower with Cu0 ENPs than Ag0 ENPs. Overall, the incorporation of ENPs into Andisols generated an increase in P retention, which may affect agricultural crop production.

Genetic Diversity and Azole Resistance Among Natural Aspergillus fumigatus Populations in Yunnan, China

2021 ◽  
Author(s):  
Duanyong Zhou ◽  
Ruirui Wang ◽  
Xiao Li ◽  
Bin Peng ◽  
Guangzhu Yang ◽  
...  
Keyword(s):  
Genetic Diversity ◽  
Aspergillus Fumigatus ◽  
Azole Resistance
Sign in / Sign up

Export Citation Format

Share Document