Allodiploid hybrid species, Aspergillus latus, belonging to section Nidulantes, is a hybrid of A. spinulosporus and an unknown species closely related to A. quadrilineatus and A. sublatus. This hybrid has often been misidentified as the species in section Nidulantes, such as A. nidulans, A. spinulosporus, A. sublatus, or other cryptic species. Aspergillus latus has not been reported in Japan as well as Asia so far. In this study, we screened 23 clinical strains identified as A. spinulosporus isolated in Japan from 2012 to 2023 and found seven A. latus strains. To characterize the A. latus strains, we conducted comprehensive phenotyping including morphological observation, whole genome sequences, and phylogenetic analysis based on calmodulin (CaM) gene. In addition, we conducted antifungal susceptibility testing for A. latus strains. As a result, the morphological characters of A. latus were more similar to those of A. spinulosporus compared to A. sublatus. However, the ascospore of A. latus differed from that of A. spinulosporus. Phylogenetic analysis revealed that different CaM alleles from the same isolate clustered separately with A. spinulosporus and A. sublatus, consistent with its hybrid origin. Furthermore, A. latus strains showed reduced susceptibility to caspofungin and amphotericin B compared to A. spinulosporus, while they were susceptible to azoles. Our results suggest that A. latus has been a causative pathogen of aspergillosis in Japan since 2013.
- MeSH
- antifungální látky farmakologie MeSH
- Aspergillus * genetika klasifikace izolace a purifikace účinky léků MeSH
- aspergilóza * mikrobiologie epidemiologie MeSH
- fylogeneze MeSH
- kalmodulin genetika MeSH
- lidé MeSH
- mikrobiální testy citlivosti MeSH
- sekvenování celého genomu MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- Geografické názvy
- Japonsko MeSH
UNLABELLED: The Aspergillus genus encompasses a diverse array of species, some of which are opportunistic pathogens. Traditionally, human aspergillosis has primarily been linked to a few Aspergillus species, predominantly A. fumigatus. Changes in epidemiology and advancements in molecular techniques have brought attention to less common and previously unrecognized pathogenic cryptic species. Despite the taxonomic recognition of many cryptic species in section Terrei, their virulence potential and clinical implications, compared to A. terreus sensu stricto, remain poorly understood. Hence, the current study utilized the alternative in vivo model Galleria mellonella to evaluate the virulence potential of 19 accepted Aspergillus species in section Terrei, classified into three series (major phylogenetic clades): Terrei, Nivei, and Ambigui. Analyzing the median survival rates of infected larvae of all species in each series revealed that series Ambigui has a significantly lower virulence compared to series Terrei and Nivei. Taking a closer look at series Terrei and Nivei revealed a trend of survival within each clade, dividing the species into two groups: highly virulent (up to 72 h survival) and less virulent (up to 144 h survival). Histological observation, considering fungal distribution and filamentation, further supported this assessment, revealing increased distribution and hyphal formation in virulent species. Additionally, the susceptibility profile of conventional antifungals was determined, revealing an increased azole minimum inhibitory concentration for some tested cryptic species such as A. niveus and A. iranicus. Our results highlight the importance of cryptic species identification, as they can exhibit different levels of virulence and show reduced antifungal susceptibility. IMPORTANCE: With changing fungal epidemiology and an increasingly vulnerable population, cryptic Aspergillus species are emerging as human pathogens. Their diversity and clinical relevance remain underexplored, with some species showing reduced antifungal susceptibility and higher virulence, highlighting the need for better preparedness in clinical practice. Using the Galleria mellonella model, we assessed the virulence of Aspergillus species of section Terrei, including cryptic and non-cryptic species, across three series Terrei, Nivei, and Ambigui. The results revealed significant virulence variation among the series, with some cryptic species displaying high virulence. Histological analysis confirmed increased hyphal formation and fungal spread in the more virulent species. Additionally, elevated azole minimum inhibitory concentrations were also observed in certain cryptic species. This study presents novel insights into the pathogenicity of Aspergillus section Terrei, emphasizing the critical importance of accurately identifying cryptic species due to their diverse virulence potential and antifungal resistance, which may have substantial clinical implications.
- MeSH
- antifungální látky farmakologie MeSH
- Aspergillus * patogenita klasifikace účinky léků genetika MeSH
- aspergilóza * mikrobiologie MeSH
- fylogeneze MeSH
- larva mikrobiologie MeSH
- lidé MeSH
- modely nemocí na zvířatech MeSH
- můry * mikrobiologie MeSH
- virulence MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- MeSH
- amfotericin B farmakologie terapeutické užití MeSH
- Aspergillus patogenita MeSH
- aspergilóza * diagnóza farmakoterapie MeSH
- debridement metody MeSH
- kazuistiky jako téma MeSH
- lidé MeSH
- mukormykóza * diagnóza farmakoterapie MeSH
- novorozenci extrémně nezralí * MeSH
- novorozenec MeSH
- Rhizopus patogenita MeSH
- Check Tag
- lidé MeSH
- novorozenec MeSH
- ženské pohlaví MeSH
- Publikační typ
- přehledy MeSH
Bacterial biofilms pose significant challenges, from healthcare-associated infections to biofouling in industrial systems, resulting in significant health impacts and financial losses globally. Classic antimicrobial methods often fail to eradicate sessile microbial communities within biofilms, requiring innovative approaches. This review explores the structure, formation, and role of biofilms, highlighting the critical importance of exopolysaccharides in biofilm stability and resistance mechanisms. We emphasize the potential of microbial enzymatic approaches, particularly focusing on glycosidases, proteases, and deoxyribonucleases, which can disrupt biofilm matrices effectively. We also delve into the importance of enzymes such as cellobiose dehydrogenase, which disrupts biofilms by degrading polysaccharides. This enzyme is mainly sourced from Aspergillus niger and Sclerotium rolfsii, with optimized production strategies enhancing its efficacy. Additionally, we explore levan hydrolase, alginate lyase, α-amylase, protease, and lysostaphin as potent antibiofilm agents, discussing their microbial origins and production optimization strategies. These enzymes offer promising avenues for combating biofilm-related challenges in healthcare, environmental, and industrial settings. Ultimately, enzymatic strategies present environmentally friendly solutions with high potential for biofilm management and infection control.
Amphotericin B (AmB) is one of the most effective antifungal drugs, with a strong, dose-dependent activity against most Candida and Aspergillus species responsible for life-threatening infections. However, AmB is severely toxic, which hinders its broad use. In this proof-of-concept study, we demonstrate that prodrugging AmB considerably decreases AmB toxicity without affecting its fungicidal activity. For this purpose, we modified the AmB structure by attaching a designer phosphate promoiety, thereby switching off its mode of action and preventing its toxic effects. The original fungicidal activity of AmB was then restored upon prodrug activation by host plasma enzymes. These AmB prodrugs showed a safer toxicity profile than commercial AmB deoxycholate in Candida and Aspergillus species and significantly prolonged larval survival of infected Galleria mellonella larvae. Based on these findings, prodrugging toxic antifungals may be a viable strategy for broadening the antifungal arsenal, opening up opportunities for targeted prodrug design.
- MeSH
- amfotericin B * farmakologie MeSH
- antifungální látky * farmakologie chemie chemická syntéza MeSH
- Aspergillus účinky léků MeSH
- Candida účinky léků MeSH
- larva účinky léků MeSH
- mikrobiální testy citlivosti * MeSH
- molekulární struktura MeSH
- můry účinky léků MeSH
- prekurzory léčiv * farmakologie chemie chemická syntéza MeSH
- vztah mezi dávkou a účinkem léčiva MeSH
- vztahy mezi strukturou a aktivitou MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
Calcineurin-nuclear factor of activated T cells (CN-NFAT) inhibitors are widely clinically used drugs for immunosuppression, but besides their required T cell response inhibition, they also undesirably affect innate immune cells. Disruption of innate immune cell function can explain the observed susceptibility of CN-NFAT inhibitor-treated patients to opportunistic fungal infections. Neutrophils play an essential role in innate immunity as a defense against pathogens; however, the effect of CN-NFAT inhibitors on neutrophil function was poorly described. Thus, we tested the response of human neutrophils to opportunistic fungal pathogens, namely Candida albicans and Aspergillus fumigatus, in the presence of CN-NFAT inhibitors. Here, we report that the NFAT pathway members were expressed in neutrophils and mediated part of the neutrophil response to pathogens. Upon pathogen exposure, neutrophils underwent profound transcriptomic changes with subsequent production of effector molecules. Importantly, genes and proteins involved in the regulation of the immune response and chemotaxis, including the chemokines CCL2, CCL3, and CCL4 were significantly upregulated. The presence of CN-NFAT inhibitors attenuated the expression of these chemokines and impaired the ability of neutrophils to chemoattract other immune cells. Our results amend knowledge about the impact of CN-NFAT inhibition in human neutrophils.
- MeSH
- Aspergillus fumigatus imunologie MeSH
- Candida albicans imunologie MeSH
- chemotaxe MeSH
- kalcineurin * metabolismus MeSH
- lidé MeSH
- mykózy imunologie MeSH
- neutrofily * imunologie metabolismus MeSH
- signální transdukce * MeSH
- transkripční faktory NFATC * metabolismus MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
Introduction. The fungal pathogen Aspergillus fumigatus can induce prolonged colonization of the lungs of susceptible patients, resulting in conditions such as allergic bronchopulmonary aspergillosis and chronic pulmonary aspergillosis.Hypothesis. Analysis of the A. fumigatus secretome released during sub-lethal infection of G. mellonella larvae may give an insight into products released during prolonged human colonisation.Methodology.Galleria mellonella larvae were infected with A. fumigatus, and the metabolism of host carbohydrate and proteins and production of fungal virulence factors were analysed. Label-free qualitative proteomic analysis was performed to identify fungal proteins in larvae at 96 hours post-infection and also to identify changes in the Galleria proteome as a result of infection.Results. Infected larvae demonstrated increasing concentrations of gliotoxin and siderophore and displayed reduced amounts of haemolymph carbohydrate and protein. Fungal proteins (399) were detected by qualitative proteomic analysis in cell-free haemolymph at 96 hours and could be categorized into seven groups, including virulence (n = 25), stress response (n = 34), DNA repair and replication (n = 39), translation (n = 22), metabolism (n = 42), released intracellular (n = 28) and cellular development and cell cycle (n = 53). Analysis of the Gallerial proteome at 96 hours post-infection revealed changes in the abundance of proteins associated with immune function, metabolism, cellular structure, insect development, transcription/translation and detoxification.Conclusion. Characterizing the impact of the fungal secretome on the host may provide an insight into how A. fumigatus damages tissue and suppresses the immune response during long-term pulmonary colonization.
- MeSH
- Aspergillus fumigatus * metabolismus MeSH
- aspergilóza mikrobiologie metabolismus MeSH
- faktory virulence metabolismus MeSH
- fungální proteiny * metabolismus genetika MeSH
- hemolymfa mikrobiologie metabolismus MeSH
- larva * mikrobiologie MeSH
- můry * mikrobiologie MeSH
- proteom analýza MeSH
- proteomika MeSH
- sekretom metabolismus MeSH
- virulence MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
CRISPR/Cas technology is a powerful tool for genome engineering in Aspergillus oryzae as an industrially important filamentous fungus. Previous study has reported the application of the CRISPR/Cpf1 system based on the Cpf1 (LbCpf1) from Lachnospiraceae bacterium in A. oryzae. However, multiplex gene editing have not been investigated using this system. Here, we presented a new CRISPR/Cpf1 multiplex gene editing system in A. oryzae, which contains the Cpf1 nuclease (FnCpf1) from Francisella tularensis subsp. novicida U112 and CRISPR-RNA expression cassette. The crRNA cassette consisted of direct repeats and guide sequences driven by the A. oryzae U6 promoter and U6 terminator. Using the constructed FnCpf1 gene editing system, the wA and pyrG genes were mutated successfully. Furthermore, simultaneous editing of wA and pyrG genes in A. oryzae was performed using two guide sequences targeting these gene loci in a single crRNA array. This promising CRISPR/Cpf1 genome-editing system provides a powerful tool for genetically engineering A. oryzae.
The pathogenic fungus Aspergillus fumigatus utilizes a cyclic ferrioxamine E (FOXE) siderophore to acquire iron from the host. Biomimetic FOXE analogues were labeled with gallium-68 for molecular imaging with PET. [68Ga]Ga(III)-FOXE analogues were internalized in A. fumigatus cells via Sit1. Uptake of [68Ga]Ga(III)-FOX 2-5, the most structurally alike analogue to FOXE, was high by both A. fumigatus and bacterial Staphylococcus aureus. However, altering the ring size provoked species-specific uptake between these two microbes: ring size shortening by one methylene unit (FOX 2-4) increased uptake by A. fumigatus compared to that by S. aureus, whereas lengthening the ring (FOX 2-6 and 3-5) had the opposite effect. These results were consistent both in vitro and in vivo, including PET imaging in infection models. Overall, this study provided valuable structural insights into the specificity of siderophore uptake and, for the first time, opened up ways for selective targeting and imaging of microbial pathogens by siderophore derivatization.
- MeSH
- Aspergillus fumigatus * metabolismus chemie MeSH
- aspergilóza * diagnostické zobrazování mikrobiologie MeSH
- biomimetické materiály chemie metabolismus MeSH
- cyklické peptidy MeSH
- deferoxamin chemie MeSH
- druhová specificita MeSH
- myši MeSH
- pozitronová emisní tomografie * metody MeSH
- radioizotopy galia * chemie MeSH
- siderofory * chemie metabolismus MeSH
- Staphylococcus aureus * metabolismus MeSH
- železité sloučeniny chemie MeSH
- zvířata MeSH
- Check Tag
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
We explored the potential of a fungal strain Aspergillus costaricensis KS1 for modulating growth and nutrient mobilization in rice. At laboratory conditions, there was a decline in pH of the medium on inoculation with the strain and the production of citric acid was observed under broth conditions. Similarly, there was higher solubilization of tricalcium phosphate and siderophore production in liquid medium on inoculation with the strain. The effect of inoculation of KS1 was studied in rice and higher growth and yield were observed on inoculation compared to control. The content of phosphorus and iron in stem and roots of KS1 inoculated plants was higher in comparison with uninoculated control. There was also increased availability of phosphorus and iron content in soil grown with KS1 inoculated plants. In addition, inoculation with strain resulted in a higher content of volatile organic compounds such as linoleic acid, linolenic acid, and ethyl isoallocholate in stem of rice. A. costaricensis KS1 can be used for improving phosphorus and iron nutrition and impart tolerance against stresses in rice.
- MeSH
- Aspergillus * metabolismus růst a vývoj MeSH
- fosfor * metabolismus analýza MeSH
- fosforečnany vápenaté metabolismus MeSH
- koncentrace vodíkových iontů MeSH
- kořeny rostlin mikrobiologie metabolismus MeSH
- kyselina citronová metabolismus MeSH
- půdní mikrobiologie MeSH
- rýže (rod) * mikrobiologie metabolismus růst a vývoj MeSH
- siderofory * metabolismus MeSH
- stonky rostlin mikrobiologie metabolismus chemie MeSH
- těkavé organické sloučeniny * metabolismus analýza MeSH
- železo * metabolismus MeSH
- Publikační typ
- časopisecké články MeSH
