Aspergillus hubkae, a Novel Species Isolated from a Patient with Probable Invasive Pulmonary Aspergillosis.

A 50-year-old man, previously diagnosed with pulmonary tuberculosis and lung cavities, presented with symptoms including fever, shortness of breath, and cough. A pulmonary CT scan revealed multiple cavities, consolidation and tree-in-bud in the upper lungs. Further investigation through direct examination of bronchoalveolar lavage fluid showed septate hyphae with dichotomous acute branching. Subsequent isolation and morphological analysis identified the fungus as belonging to Aspergillus section Nigri. The patient was diagnosed with probable invasive pulmonary aspergillosis and successfully treated with a three-month oral voriconazole therapy. Phylogenetic analysis based on partial ß-tubulin, calmodulin and RNA polymerase second largest subunit sequences revealed that the isolate represents a putative new species related to Aspergillus brasiliensis, and is named Aspergillus hubkae here. Antifungal susceptibility testing demonstrated that the isolate is resistant to itraconazole but susceptible to voriconazole. This phenotypic and genetic characterization of A. hubkae, along with the associated case report, will serve as a valuable resource for future diagnoses of infections caused by this species. It will also contribute to more precise and effective patient management strategies in similar clinical scenarios.

Enhancement of soil aggregation and physical properties through fungal amendments under varying moisture conditions.

Soil structure and aggregation are crucial for soil functionality, particularly under drought conditions. Saprobic soil fungi, known for their resilience in low moisture conditions, are recognized for their influence on soil aggregate dynamics. In this study, we explored the potential of fungal amendments to enhance soil aggregation and hydrological properties across different moisture regimes. We used a selection of 29 fungal isolates, recovered from soils treated under drought conditions and varying in colony density and growth rate, for single-strain inoculation into sterilized soil microcosms under either low or high moisture (≤-0.96 and -0.03 MPa, respectively). After 8 weeks, we assessed soil aggregate formation and stability, along with soil properties such as soil water content, water hydrophobicity, sorptivity, total fungal biomass and water potential. Our findings indicate that fungal inoculation altered soil hydrological properties and improved soil aggregation, with effects varying based on the fungal strains and soil moisture levels. We found a positive correlation between fungal biomass and enhanced soil aggregate formation and stabilization, achieved by connecting soil particles via hyphae and modifying soil aggregate sorptivity. The improvement in soil water potential was observed only when the initial moisture level was not critical for fungal activity. Overall, our results highlight the potential of using fungal inoculation to improve the structure of agricultural soil under drought conditions, thereby introducing new possibilities for soil management in the context of climate change.

Blastobotrys nigripullensis, a new yeast species isolated from a fungal outbreak on an ancient Roman shipwreck in the Netherlands.

A new species of the yeast genus Blastobotrys was discovered on ancient ship timbers in the Netherlands. The species had developed on the wood of a river barge dating to the Roman period. The growth occurred after the preservative polyethylene glycol (PEG 4000) was washed out of some of the timbers due to an undetected leak in the storage unit. Mycological analysis of various timber samples revealed the presence of Microascus melanosporus (predominant), Microascus paisii, a member of the Acremonium chrysogenum-clade, and a new Blastrobotrys species. The new species produced sporothrix-like conidiophores with clavate blastoconidia (3-7 × 1-3.5 µm) and was found to be osmotolerant, capable of growth on low water activity media like malt yeast 50% glucose agar (MY50G). In this article we formally describe and introduce Blastrobotrys nigripullensis (CBS 17879 T) based on its morphology, physiology and phylogenetic placement.

Regional distribution and diversity of Aspergillus and Penicillium species on Croatian traditional meat products.

Various factors, such as weather and production practices (e.g., environmental hygiene, process duration, raw material quality, ripening temperature, and relative humidity), in combination with the intrinsic product properties (e.g., pH, aw, salt content), significantly affect the growth of surface moulds. The aim of this study was to isolate and identify surface moulds retrieved from traditional meat products (TMPs) and correlate these data to the production region and production technology. The surface of 250 TMPs (dry-fermented sausages, n = 108; dry-cured meat products, n = 142) from five Croatian regions were sampled during a two-year period. Dry-fermented sausages had a significantly higher pH and a lower salt concentration when compared to dry-cured meat products. In total, 528 isolates were obtained, comprising 20 Penicillium and 17 Aspergillus species. The species most frequently isolated from the dry-fermented sausages were P. commune (32.4 %), A. proliferans (33 %), and P. solitum (14.8 %), while A. proliferans (52.1 %), P. commune (28.9 %) and P. citrinum (19.7 %) predominated in dry-cured meat products. Aspergillus predominated on the TMPs from southern Croatia, while Penicillium was prevalent on products from the other four regions, possibly due to differences in weather conditions. Seven potentially mycotoxigenic species (A. creber, A. flavus, A. niger, A. westerdijkiae, P. citrinum, P. commune, and P. nordicum) were isolated and identified. Regular monitoring of mould species and their toxigenic metabolites present on traditional meat products is of the utmost importance from the public health perspective, while the results of such a monitoring can prove beneficial for the tailoring of the production technology development.

Fungal Diversity and Aflatoxins in Maize and Rice Grains and Cassava-Based Flour (Pupuru) from Ondo State, Nigeria.

Grains and cassava-based foods serve as major dietary sources for many households in Nigeria. However, these foods are highly prone to contamination by moulds and aflatoxins owing to poor storage and vending practices. Therefore, we studied the fungal diversity in maize, cassava-based flour (pupuru), and rice vended in markets from Ondo state, Nigeria, and assessed their aflatoxin levels using an enzyme-linked immunosorbent assay. Molecular analysis of 65 representative fungal isolates recovered from the ground grains and pupuru samples revealed 26 species belonging to five genera: Aspergillus (80.9%), Penicillium (15.4%), and Talaromyces (1.9%) in the Ascomycota; Syncephalastrum (1.2%) and Lichtheimia (0.6%) in Mucoromycota. Aspergillus flavus was the predominant species in the ground grains and pupuru samples. Aflatoxins were found in 73.8% of the 42 representative food samples and 41.9% exceeded the 10 µg/kg threshold adopted in Nigeria for total aflatoxins.

The Environmental Spread of Aspergillus terreus in Tyrol, Austria.

Fungal infections due to Aspergillus species have become a major cause of morbidity and mortality among immunocompromised patients. At the Medical University of Innsbruck, A. terreus and related species are the second most common causative agents of aspergillosis. In this one-year study we collected environmental samples to investigate (i) the environmental distribution, (ii) the ecological niche of A. terreus in Tyrol, (iii) the genetic relatedness of environmental and clinical isolates and the correlation between those two groups of isolates, and (iv) the antifungal susceptibility patterns. A. terreus was present in 5.4% of 3845 environmental samples, with a significantly higher frequency during winter (6.8%) than summer (3.9%). An increased A. terreus abundance in Tyrol's Eastern part was detected which is in agreement with the proof of clinical cases. In total, 92% of environmental and 98% of clinical A. terreus isolates were amphotericin B resistant; 22.6% and 9.8% were resistant against posaconazole. Overall, 3.9% of clinical isolates were resistant against voriconazole. Short tandem repeat analysis identified three major genotypes persisting in Tyrol. Soil from agricultural cornfields seems to be an important source; the environmental frequency of A. terreus correlates with the high incidence of A. terreus infections in certain geographical areas.

Diversity and toxigenicity of fungi and description of Fusarium madaense sp. nov. from cereals, legumes and soils in north-central Nigeria.

Mycological investigation of various foods (mainly cowpea, groundnut, maize, rice, sorghum) and agricultural soils from two states in north-central Nigeria (Nasarawa and Niger), was conducted in order to understand the role of filamentous fungi in food contamination and public health. A total of 839 fungal isolates were recovered from 84% of the 250 food and all 30 soil samples. Preliminary identifications were made, based on macro- and micromorphological characters. Representative strains (n = 121) were studied in detail using morphology and DNA sequencing, involving genera/species-specific markers, while extrolite profiles using LC-MS/MS were obtained for a selection of strains. The representative strains grouped in seven genera (Aspergillus, Fusarium, Macrophomina, Meyerozyma, Neocosmospora, Neotestudina and Phoma). Amongst the 21 species that were isolated during this study was one novel species belonging to the Fusarium fujikuroi species complex, F. madaense sp. nov., obtained from groundnut and sorghum in Nasarawa state. The examined strains produced diverse extrolites, including several uncommon compounds: averantinmethylether in A. aflatoxiformans; aspergillimide in A. flavus; heptelidic acid in A. austwickii; desoxypaxillin, kotanin A and paspalitrems (A and B) in A. aflatoxiformans, A. austwickii and A. cerealis; aurasperon C, dimethylsulochrin, fellutanine A, methylorsellinic acid, nigragillin and pyrophen in A. brunneoviolaceus; cyclosporins (A, B, C and H) in A. niger; methylorsellinic acid, pyrophen and secalonic acid in A. piperis; aspulvinone E, fonsecin, kojic acid, kotanin A, malformin C, pyranonigrin and pyrophen in A. vadensis; and all compounds in F. madaense sp. nov., Meyerozyma, Neocosmospora and Neotestudina. This study provides snapshot data for prediction of food contamination and fungal biodiversity exploitation.

Thermotolerant and Thermophilic Mycobiota in Different Steps of Compost Maturation.

Composting is a complex process in which various micro-organisms, mainly fungi and bacteria, are involved. The process depends on a large number of factors (biological, chemical, and physical) among which microbial populations play a fundamental role. The high temperatures that occur during the composting process indicate the presence of thermotolerant and thermophilic micro-organisms that are key for the optimization of the process. However, the same micro-organisms can be harmful (allergenic, pathogenic) for workers that handle large quantities of material in the plant, and for end users, for example, in the indoor environment (e.g., pots in houses and offices). Accurate knowledge of thermotolerant and thermophilic organisms present during the composting stages is required to find key organisms to improve the process and estimate potential health risks. The objective of the present work was to study thermotolerant and thermophilic mycobiota at different time points of compost maturation. Fungi were isolated at four temperatures (25, 37, 45, and 50 °C) from compost samples collected at five different steps during a 21-day compost-maturation period in an active composting plant in Liguria (northwestern Italy). The samples were subsequently plated on three different media. Our results showed a high presence of fungi with an order of magnitude ranging from 1 × 104 to 3 × 105 colony-forming units (CFU) g-1. The isolated strains, identified by means of specific molecular tools (ITS, beta-tubulin, calmodulin, elongation factor 1-alpha, and LSU sequencing), belonged to 45 different species. Several thermophilic species belonging to genera Thermoascus and Thermomyces were detected, which could be key during composting. Moreover, the presence of several potentially harmful fungal species, such as Aspergillus fumigatus, A. terreus, and Scedosporium apiospermum, were found during the whole process, including the final product. Results highlighted the importance of surveying the mycobiota involved in the composting process in order to: (i) find solutions to improve efficiency and (ii) reduce health risks.

Fungal Diversity and Mycotoxins in Low Moisture Content Ready-To-Eat Foods in Nigeria.

Low moisture content ready-to-eat foods vended in Nigerian markets could be pre-packaged or packaged at point of sale. These foods are widely and frequently consumed across Nigeria as quick foods. Despite their importance in the daily diets of Nigerians, a comprehensive study on the diversity of fungi, fungal metabolite production potential, and mycotoxin contamination in the foods has not yet been reported. Therefore, this study assessed the diversity of fungi in 70 samples of low moisture content ready-to-eat foods [cheese balls, garri (cassava-based), granola (a mix of cereals and nuts) and popcorn] in Nigeria by applying a polyphasic approach including morphological examination, genera/species-specific gene marker sequencing and secondary metabolite profiling of fungal cultures. Additionally, mycotoxin levels in the foods were determined by LC-MS/MS. Fungal strains (n = 148) were recovered only from garri. Molecular analysis of 107 representative isolates revealed 27 species belonging to 12 genera: Acremonium, Allophoma, Aspergillus, Cladosporium, Fusarium, Microdochium, Penicillium, Sarocladium, Talaromyces, and Tolypocladium in the Ascomycota, and Fomitopsis and Trametes in the Basidiomycota. To the best of our knowledge Allophoma, Fomitopsis, Microdochium, Tolypocladium, and Trametes are reported in African food for the first time. A total of 21 uncommon metabolites were found in cultures of the following species: andrastin A and sporogen AO1 in Aspergillus flavus; paspalin in A. brunneoviolaceus; lecanoic acid and rugulusovin in A. sydowii; sclerotin A in P. citrinum and Talaromyces siamensis; barceloneic acid, festuclavine, fumigaclavine, isochromophilons (IV, VI, and IX), ochrephilone, sclerotioramin, and sclerotiorin in P. sclerotium; epoxyagroclavine, infectopyron, methylorsellinic acid and trichodermamide C in P. steckii; moniliformin and sporogen AO1 in P. copticola; and aminodimethyloctadecanol in Tolypocladium. Twenty-four mycotoxins in addition to other 73 fungal and plant toxins were quantified in the foods. In garri, cheeseballs, popcorn and granola were 1, 6, 12, and 23 mycotoxins detected, respectively. Deoxynivalenol, fumonisins, moniliformin, aflatoxins and citrinin contaminated 37, 31, 31, 20, and 14% of all food samples, respectively. Overall, citrinin had the highest mean concentration of 1481 µg/kg in the foods, suggesting high citrinin exposures in the Nigerian populace. Fungal and mycotoxin contamination of the foods depend on pre-food and post-food processing practices.

Cathelicidin-inspired antimicrobial peptides as novel antifungal compounds.

Fungal infections in humans are increasing worldwide and are currently mostly treated with a relative limited set of antifungals. Resistance to antifungals is increasing, for example, in Aspergillus fumigatus and Candida auris, and expected to increase for many medically relevant fungal species in the near future. We have developed and patented a set of cathelicidin-inspired antimicrobial peptides termed 'PepBiotics'. These peptides were initially selected for their bactericidal activity against clinically relevant Pseudomonas aeruginosa and Staphylococcus aureus isolates derived from patients with cystic fibrosis and are active against a wide range of bacteria (ESKAPE pathogens). We now report results from studies that were designed to investigate the antifungal activity of PepBiotics against a set of medically relevant species encompassing species of Aspergillus, Candida, Cryptococcus, Fusarium, Malassezia, and Talaromyces. We characterized a subset of PepBiotics and show that these peptides strongly affected metabolic activity and/or growth of a set of medically relevant fungal species, including azole-resistant A. fumigatus isolates. PepBiotics showed a strong inhibitory activity against a large variety of filamentous fungi and yeasts species at low concentrations (≤1 µM) and were fungicidal for at least a subset of these fungal species. Interestingly, the concentration of PepBiotics required to interfere with growth or metabolic activity varied between different fungal species or even between isolates of the same fungal species. This study shows that PepBiotics display strong potential for use as novel antifungal compounds to fight a large variety of clinically relevant fungal species.

Genomic insights into the atopic eczema-associated skin commensal yeast Malassezia sympodialis.

UNLABELLED: Malassezia commensal yeasts are associated with a number of skin disorders, such as atopic eczema/dermatitis and dandruff, and they also can cause systemic infections. Here we describe the 7.67-Mbp genome of Malassezia sympodialis, a species associated with atopic eczema, and contrast its genome repertoire with that of Malassezia globosa, associated with dandruff, as well as those of other closely related fungi. Ninety percent of the predicted M. sympodialis protein coding genes were experimentally verified by mass spectrometry at the protein level. We identified a relatively limited number of genes related to lipid biosynthesis, and both species lack the fatty acid synthase gene, in line with the known requirement of these yeasts to assimilate lipids from the host. Malassezia species do not appear to have many cell wall-localized glycosylphosphatidylinositol (GPI) proteins and lack other cell wall proteins previously identified in other fungi. This is surprising given that in other fungi these proteins have been shown to mediate interactions (e.g., adhesion and biofilm formation) with the host. The genome revealed a complex evolutionary history for an allergen of unknown function, Mala s 7, shown to be encoded by a member of an amplified gene family of secreted proteins. Based on genetic and biochemical studies with the basidiomycete human fungal pathogen Cryptococcus neoformans, we characterized the allergen Mala s 6 as the cytoplasmic cyclophilin A. We further present evidence that M. sympodialis may have the capacity to undergo sexual reproduction and present a model for a pseudobipolar mating system that allows limited recombination between two linked MAT loci. IMPORTANCE: Malassezia commensal yeasts are associated with a number of skin disorders. The previously published genome of M. globosa provided some of the first insights into Malassezia biology and its involvement in dandruff. Here, we present the genome of M. sympodialis, frequently isolated from patients with atopic eczema and healthy individuals. We combined comparative genomics with sequencing and functional characterization of specific genes in a population of clinical isolates and in closely related model systems. Our analyses provide insights into the evolution of allergens related to atopic eczema and the evolutionary trajectory of the machinery for sexual reproduction and meiosis. We hypothesize that M. sympodialis may undergo sexual reproduction, which has important implications for the understanding of the life cycle and virulence potential of this medically important yeast. Our findings provide a foundation for the development of genetic and genomic tools to elucidate host-microbe interactions that occur on the skin and to identify potential therapeutic targets.