Portuguese cork industry: filling the knowledge gap regarding occupational exposure to fungi and related health effects.

Introduction: The presence of the Penicillium section Aspergilloides (formerly known as Penicillium glabrum) in the cork industry involves the risk of respiratory diseases such as suberosis. Methods: The aim of this study was to corroborate the predominant fungi present in this occupational environment by performing a mycological analysis of 360 workers' nasal exudates collected by nasal swabs. Additionally, evaluation of respiratory disorders among the cork workers was also performed by spirometry. Results: Penicillium section Aspergilloides was detected by qPCR in 37 out of the 360 nasal swabs collected from workers' samples. From those, 25 remained negative for Penicillium sp. when using culture-based methods. A significant association was found between ventilatory defects and years of work in the cork industry, with those people working for 10 or more years in this industry having an approximately two-fold increased risk of having ventilatory defects compared to those working less time in this setting. Among the workers who detected the presence of Penicillium section Aspergilloides, those with symptoms presented slightly higher average values of CFU. Discussion: Overall, the results obtained in this study show that working in the cork industry may have adverse effects on worker's respiratory health. Nevertheless, more studies are needed (e.g., using serological assays) to clarify the impact of each risk factor (fungi and dust) on disease etiology.

Secondary Metabolites with Agricultural Antagonistic Potential from Aspergillus sp. ITBBc1, a Coral-Associated Marine Fungus.

A marine-derived fungal strain, Aspergillus sp. ITBBc1, was isolated from coral collected from the South China Sea in Hainan province. Intensive chemical investigation of the fermentation extract of this strain afforded four new secondary metabolites (1-4), named megastigmanones A-C and prenylterphenyllin H, along with four known compounds (5-8). Their structures were elucidated by extensive spectroscopic analysis including one-and two-dimensional (1D and 2D) NMR spectroscopy and high-resolution electrospray ionization mass spectrometry (HR-ESI-MS). The modified Mosher's method was undertaken to determine the absolute configurations of new compounds. The phytotoxic activity test showed that compounds 6-8 exhibited significant antagonistic activity against the germination of Triticum aestivum L. and Oryza sativa L. seeds with a dose-dependent relationship.

New Meroterpenoids and α-Pyrone Derivatives Isolated from the Mangrove Endophytic Fungal Strain Aspergillus sp. GXNU-Y85.

Two new meroterpenoids, aspergienynes O and P (1 and 2), one new natural compound, aspergienyne Q (3), and a new α-pyrone derivative named 3-(4-methoxy-2-oxo-2H-pyran-6-yl)butanoic acid (4) were isolated from the mangrove endophytic fungal strain Aspergillus sp. GXNU-Y85, along with five known compounds (5-9). The absolute configurations of those new isolates were confirmed through extensive analysis using spectroscopic data (HRESIMS, NMR, and ECD). The pharmacological study of the anti-proliferation activity indicated that isolates 5 and 9 displayed moderate inhibitory effects against HeLa and A549 cells, with the IC50 values ranging from 16.6 to 45.4 µM.

Multifunctional hydrophobin-like protein (HFB-NJ1): A versatile solution for wastewater treatment.

As wastewater contains a variety of contaminating bacteria and oily residues, there is an urgent need for environmentally safe bactericidal agents and surfactants which can be applied for wastewater treatment. The present study emphasizes on the potential of hydrophobin-like protein (HFB-NJ1) extracted from sporulating mycelia of Aspergillus sp. NJ1 for wastewater treatment. The purified HFB-NJ1, depicted the presence of one single protein band of molecular size approximately 11-12 kDa on silver-stained SDS-PAGE gel. HFB-NJ1 also presented properties such as surface modification of glass and stable emulsification of sunflower oil. HFB-NJ1 depicted exceptional antibacterial activity against bacterial pathogens such as Bacillus subtilis and Pseudomonas aeruginosa at low MIC of 0.5 µg/mL and 0.75 µg/mL respectively. Additionally, HFB-NJ1 depicted enhanced emulsification of various vegetable and petroleum-based oils (E24 > 80%). HFB-NJ1 effectively reduced gold ions, producing nanospheres with a size of 15.33 nm - a recognized antimicrobial agent. This study underscores the multifunctional attributes of HFB-NJ1, highlighting its efficacy in removing pathogenic bacteria, emulsifying organic compounds from wastewater, and demonstrating a reduction ability for nanoparticle synthesis.

Bio-fabricated zinc oxide nanoparticles mediated by endophytic fungus Aspergillus sp. SA17 with antimicrobial and anticancer activities: in vitro supported by in silico studies.

Introduction: In recent years, the world's attention has been drawn to antimicrobial resistance (AMR) because to the frightening prospect of growing death rates. Nanomaterials are being investigated due to their potential in a wide range of technical and biological applications. Methods: The purpose of this study was to biosynthesis zinc oxide nanoparticles (ZnONPs) using Aspergillus sp. SA17 fungal extract, followed by characterization of the produced nanoparticles (NP) using electron microscopy (TEM and SEM), UV-analysis, X-ray diffraction (XRD), and Fourier-transform infrared spectroscopy (FT-IR). Results and Discussion: The HR-TEM revealed spherical nanoparticles with an average size of 7.2 nm, and XRD validated the crystalline nature and crystal structure features of the generated ZnONPs, while the zeta potential was 18.16 mV, indicating that the particles' surfaces are positively charged. The FT-IR was also used to identify the biomolecules involved in the synthesis of ZnONPs. The antibacterial and anticancer properties of both the crude fungal extract and its nano-form against several microbial strains and cancer cell lines were also investigated. Inhibition zone diameters against pathogenic bacteria ranged from 3 to 13 mm, while IC50 values against cancer cell lines ranged from 17.65 to 84.55 M. Additionally, 33 compounds, including flavonoids, phenolic acids, coumarins, organic acids, anthraquinones, and lignans, were discovered through chemical profiling of the extract using UPLC-QTOF-MS/MS. Some molecules, such pomiferin and glabrol, may be useful for antibacterial purposes, according to in silico study, while daidzein 4'-sulfate showed promise as an anti-cancer metabolite.

Antimicrobial metabolites from the marine-derived fungus Aspergillus sp. ZZ1861.

Fungi from the genus Aspergillus are important resources for the discovery of bioactive agents. This investigation characterized the isolation, structural elucidation, and antimicrobial evaluation of 46 metabolites produced by the marine-derived fungus Aspergillus sp. ZZ1861 in rice solid and potato dextrose broth liquid media. The structures of these isolated compounds were determined based on their HRESIMS data, NMR spectral analyses, and data from ECD, NMR, and optical rotation calculations. Emericelactones F and G, 20R,25S-preshamixanthone, 20R,25R-preshamixanthone, phthalimidinic acid A, phthalimidinic acid B, aspergilol G, and 2-hydroxyemodic amide are eight previously undescribed compounds and (S)-2-(5-hydroxymethyl-2-formylpyrrol-1-yl) propionic acid lactone is reported from a natural resource for the first time. It is also the first report of the configurations of 25S-O-methylarugosin A, 25R-O-methylarugosin A, 5R-(+)-9-hydroxymicroperfuranone, and 5R-(+)-microperfuranone. Phthalimidinic acid A, phthalimidinic acid B, aspergilol G, and 2-hydroxyemodic amide have antifungal activity against Candida albicans with MIC values of 1.56, 3.12, 1.56, and 12.5 µg/mL, respectively, 20R,25S-preshamixanthone (MIC 25 µg/mL) shows antibacterial activity against Escherichia coli, and 20R,25R-preshamixanthone exhibits antimicrobial activity against all three tested pathogens of methicillin-resistant Staphylococcus aureus, E. coli, and C. albicans with MIC values of 50, 25, 25 µg/mL, respectively.

Re-discovery of MS-347a as a fungicide candidate through a new drug discovery platform with a multidrug-sensitive Saccharomyces cerevisiae screening system and the introduction of a global secondary metabolism regulator, laeA gene.

We found that the culture broth of fungi showed anti-fungal activity against multidrug-sensitive budding yeast. However, we could not identify the anti-fungal compound due to the small quantity. Therefore, we attempted to increase the productivity of the target compound by the introduction of a global secondary metabolism regulator, laeA to the strain, which led to the successful isolation of 10-folds greater amount of MS-347a (1) than Aspergillus sp. FKI-5362. Compound 1 was not effective against Candida albicans and the detailed anti-fungal activity of 1 remains unverified. After our anti-fungal activity screening, 1 was found to inhibit the growth of broad plant pathogenic fungal species belonging to the Ascomycota. It is noteworthy that 1 showed little insecticidal activity against silkworms, suggesting its selective biological activity against plant pathogenic fungi. Our study implies that the combination strategy of multidrug-sensitive yeast and the introduction of laeA is useful for new anti-fungal drug discovery.

Safety evaluation of the food enzyme AMP deaminase from non-genetically modified Aspergillus sp. strain DEA 56-111.

The food enzyme AMP deaminase (AMP aminohydrolase; EC 3.5.4.6) is produced with the non-genetically modified microorganism Aspergillus sp. strain DEA 56-111 by Shin Nihon Chemical Co., Ltd. The food enzyme was considered free from viable cells of the production organism. It is intended to be used in the processing of yeast and yeast products. Dietary exposure to the food enzyme-total organic solids (TOS) was estimated to be up to 0.005 mg TOS/kg body weight (bw) per day in European populations. Genotoxicity tests did not indicate a safety concern. The Panel identified a no observed adverse effect level of 1984 mg TOS/kg bw per day, the highest dose tested, which, when compared with the estimated dietary exposure, resulted in a margin of exposure of at least 396,800. A search for the similarity of the amino acid sequence of the food enzyme to known allergens was made and no match was found. The Panel considered that the risk of allergic reactions upon dietary exposure cannot be excluded, but the likelihood is low. Based on the data provided, the Panel concluded that this food enzyme does not give rise to safety concerns, under the intended conditions of use.

Assessment of Portuguese fitness centers: Bridging the knowledge gap on harmful microbial contamination with focus on fungi.

The lack of knowledge regarding the extent of microbial contamination in Portuguese fitness centers (FC) puts attendees and athletes at risk for bioaerosol exposure. This study intends to characterize microbial contamination in Portuguese FC by passive sampling methods: electrostatic dust collectors (EDC) (N = 39), settled dust (N = 8), vacuum filters (N = 8), and used cleaning mops (N = 12). The obtained extracts were plated in selective culture media for fungi and bacteria. Filters, EDC, and mop samples' extracts were also screened for antifungal resistance and used for the molecular detection of the selected Aspergillus sections. The detection of mycotoxins was conducted using a high-performance liquid chromatograph (HPLC) system and to determine the cytotoxicity of microbial contaminants recovered by passive sampling, HepG2 (human liver carcinoma) and A549 (human alveolar epithelial) cells were employed. The results reinforce the use of passive sampling methods to identify the most critical areas and identify environmental factors that influence microbial contamination, namely having a swimming pool. The cardio fitness area presented the highest median value of total bacteria (TSA: 9.69 × 102 CFU m-2.day-1) and Gram-negative bacteria (VRBA: 1.23 CFU m-2.day-1), while for fungi it was the open space area, with 1.86 × 101 CFU m-2.day-1. Aspergillus sp. was present in EDC and in filters used to collect settled dust. Reduced azole susceptibility was observed in filters and EDC (on ICZ and VCZ), and in mops (on ICZ). Fumonisin B2 was the only mycotoxin detected and it was present in all sampling matrixes except settled dust. High and moderate cytotoxicity was obtained, suggesting that A549 cells were more sensitive to samples' contaminants. The observed widespread of critical toxigenic fungal species with clinical relevance, such as Aspergillus section Fumigati, as well as Fumonisin B2 emphasizes the importance of frequent and effective cleaning procedures while using shared mops appeared as a vehicle of cross-contamination.

Design, semi-synthesis of soft coral-derived Aspergillus sp. secondary metabolite geodin derivatives and their antibacterial activities.

A series of novel ester derivatives 2 - 7, of natural product geodin 1, isolated from the soft coral-derived fungus Aspergillus sp., were designed and semi-synthesised through one step reaction with high yield. Compound 5 showed strong antifouling inhibitory activities with MIC of 4.80 µM while compound 4 showed selective inhibitory activities with MICs values 8.59 µM against Aeromonas salmonicida and Pseudomonas aeruginosa (Sea-Nine 211, MIC = 0.27 µM). Compounds 3, 4 and 6 showed potent anti-pathogenic inhibitory activities with MICs of 2.29 µM, 4.29 µM and 4.56 µM respectively against Staphylococcus aureus (Ciprofloxacin, MIC = 0.156 µM). Compound 2 showed weak inhibitory activity against A. salmonicida with MIC 18.75 µM (Sea-Nine 211, MIC = 0.27 µM) and with MICs 9.38 µM against S. aureus (ciprofloxacin, MIC = 0.156 µM). However, compound 7 showed very low antibacterial activities with MIC = >20 µM. The preliminary structure-activity relationships of compounds 2 - 7 further prove that the modification of 4-OH group of natural product geodin 1 improves the antibacterial activities such as antifouling and anti-pathogenic activities.

Impact of Aspergillus fumigatus inoculation on the composting of wood shaving bedding for horses.

Equine farming generates a significant amount of waste, prompting the need for effective management. Composting enhanced by filamentous fungi holds promise for this purpose. This study focused on inoculating Aspergillus fumigatus isolates in composting horse bedding made with wood shavings (Pinus elliottii). The experiment lasted 90 days, with two treatment groups, control and inoculated, analyzing temperature, pH, electrical conductivity, total organic carbon and nitrogen content, and cellulose, hemicellulose, and lignin contents. Both treatments entered the thermophilic phase by the fourth day, reaching temperatures above 55°C and mesophilic maturation at 35 days (41 ± 0.2°C). The inoculated treatment exhibited higher electrical conductivity after 30 days and a more pronounced reduction in the total carbon content (42.85% vs. 38.29%) compared to the control. While there was no significant nitrogen difference, the inoculated treatment had a sharper reduction in carbon/nitrogen ratio, and cellulose and hemicellulose contents. Both treatments showed low coliform counts, no Salmonella sp., and reduced Strongyloides sp. larvae. Inoculating A. fumigatus in saturated horse bedding made from wood shavings improved compost quality, providing a possibility for sustainable equine farming waste treatment.

Isolation, identification, and molecular characterization of potential keratinolytic fungus sp. from Southern Assam: relevance to poultry wastes and its biological management.

Feather waste is a highly prevalent form of keratinous waste that is generated by the poultry industry. The global daily production of feather waste has been shown to approach 5 million tons, typically being disposed of through methods such as dumping, landfilling, or incineration which contribute significantly to environmental pollutions. The proper management of these keratinous wastes is crucial to avoid environmental contamination. The study was carried out to isolate the keratinolytic fungi from the poultry disposal sites of different region of North-East India to evaluate its potential in bioremediation of the feathers wastes. Out of 12 fungal strains isolated from the sites, the fungus showing the highest zone of hydrolysis on both the skim milk and keratin agar medium was selected for the study and the molecular identification of the isolate was performed through DNA sequence analysis by amplifying the internal transcribed spacer (ITS) region. The sequence results showed higher similarity (above 95%) with Aspergillus spp. and was named Aspergillus sp. Iro-1. The strain was further analyzed for its feather degrading potential which was performed in submerged conditions under optimized conditions. The study showed that the strain could effectively degrade the feathers validated through weight loss method, and the structural deformations in the feathers were visualized through scanning electron microscopy (SEM). Aspergillus sp. Iro-1 was obtained from the southern region of Assam. It would be of great importance as the implementation of this sp. can help in the bioremediation of feathers wastes in this region. This is the first study of identification of feather degrading fungus from southern part of Assam (Barak).

Optimization of Fermentation Process for New Anti-Inflammatory Glycosylceramide Metabolite from Aspergillus sp.

A novel ceramide compound, named Aspercerebroside A (AcA), was successfully isolated from the ethyl acetate layer of the marine symbiotic fungus Aspergillus sp. AcA exhibited notable anti-inflammatory activity by effectively inhibiting the production of nitric oxide (NO) in RAW 264.7 cells at concentrations of 30 µg/mL and 40 µg/mL, offering a promising avenue for the treatment of inflammatory diseases. To optimize the yield of glycosylceramide (AcA), a series of techniques, including single-factor experiments, orthogonal experiments, and response surface optimization, were systematically employed to fine-tune the composition of the fermentation medium. Initially, the optimal carbon source (sucrose), nitrogen source (yeast extract powder), and the most suitable medium salinity (14 ppt) were identified through single-factor experiments. Subsequently, orthogonal experiments, employing an orthogonal table for planning and analyzing multifactor experiments, were conducted. Finally, a mathematical model, established using a Box-Behnken design, comprehensively analyzed the interactions between the various factors to determine the optimal composition of the fermentation medium. According to the model's prediction, when the sucrose concentration was set at 37.47 g/L, yeast extract powder concentration at 19.66 g/L, and medium salinity at 13.31 ppt, the predicted concentration of glycosylceramide was 171.084 µg/mL. The experimental results confirmed the model's accuracy, with the actual average concentration of glycosylceramide under these conditions measured at 171.670 µg/mL, aligning closely with the predicted value.

Diisoprenyl Cyclohexene-Type Meroterpenoids with Cytotoxic Activity from a Mangrove Endophytic Fungus Aspergillus sp. GXNU-Y85.

Five new diisoprenyl cyclohexene-type meroterpenoids, aspergienynes J-N (1-5), along with three known analogues (6-8), were obtained from the mangrove endophytic fungal strain Aspergillus sp. GXNU-Y85. The chemical structures, including their absolute configurations, were established via spectroscopic data and comparison of experimental and calculated ECD spectra. Cytotoxicity assay results indicated that compound 8 had strong cytotoxicity against HeLa cancer cells, and its IC50 value was 11.8 µM. In addition, flow cytometry analysis revealed that the cytotoxicity of 8 was due to the induction of G1 cell cycle arrest and apoptosis in HeLa cells.

Safety evaluation of the food enzyme leucyl aminopeptidase from the non-genetically modified Aspergillus sp. strain AE-MB.

The food enzyme leucyl aminopeptidase (EC 3.4.11.1) is produced with the non-genetically modified Aspergillus sp. strain AE-MB by Amano Enzyme Inc. The food enzyme is considered free from viable cells of the production organism. It is intended to be used in five food manufacturing processes: processing of dairy products for the production of (1) flavouring preparations; processing of plant- and fungal-derived products for the production of (2) protein hydrolysates; processing of meat and fish products for the production of (3) protein hydrolysates, (4) modified meat and fish products and processing of (5) yeast and yeast products. Dietary exposure to the food enzyme-total organic solids (TOS) was estimated to be up to 2.273 mg TOS/kg body weight (bw) per day in European populations. Genotoxicity tests did not indicate a safety concern. The systemic toxicity was assessed by means of a repeated dose 90-day oral toxicity study in rats. The Panel identified a no observed adverse effect level of 183 mg TOS/kg bw per day. The calculated margin of exposure for each age group was 135 (infants), 81 (toddlers), 83 (children), 109 (adolescents), 160 (adults) and 184 (the elderly). A search for the similarity of the amino acid sequence of the food enzyme to known allergens was made and no matches were found. The Panel considered that the risk of allergic reactions by dietary exposure cannot be excluded, but the likelihood is low. The safety of the food enzyme could not be established given the derived margins of exposure. Therefore, the Panel concluded that this food enzyme could not be considered safe under the intended conditions of use.

Cytotoxic metabolites from the mangrove endophytic fungus Aspergillus sp. GXNU QG1a.

A new benzoquinone, guxiumasperone A (1), and a new diisoprenyl-cyclohexene-type meroterpenoids, biscognienyne M (2), along with four known diisoprenyl-cyclohexene analogues was isolated from the mangrove endophytic fungus Aspergillus QG1a. Their structures were determined by extensive spectral analysis of 1D and 2D NMR, HR-ESI- MS, and X-ray crystallography. Compound 1 was deduced by a single-crystal X-ray diffraction analysis, and the absolute configuration of 2 was further unequivocally elucidated by comparing the experimental electronic circular dichroism (ECD) data with calculated ECD spectra. Compounds 1 and 2 showed significant cytotoxic activity against selected tumour cells. Particularly, compound 2 exhibited strong activity against A2780 cancer cells with an IC50 value of 6.8 µM.

Impact of Ascaris lumbricoides infection on the development of chronic pulmonary aspergillosis in patients with COPD.

The aetiopathogenesis of chronic obstructive pulmonary disease (COPD) remains unclear. The aim of our study was to determine the possible influence of Ascaris lumbricoides on the development of chronic pulmonary aspergillosis (CPA) in patients with COPD. The prevalence of A. lumbricoides in patients with COPD with CPA (19.05%) was significantly higher than that in those without (9.20%) and controls (4.9%) (p < 0.05). Trends in levels of Interleukin-1ß and of tumour necrosis factor α suggest ascariasis increases susceptibility to Aspergillus sp. in patients with COPD and can be considered an additional risk factor for CPA.

Isolation and Structure Elucidation of New Metabolites from the Mariana-Trench-Associated Fungus Aspergillus sp. SY2601.

Fungi are important resource for the discovery of novel bioactive natural products. This study investigated the metabolites produced by Mariana-Trench-associated fungus Aspergillus sp. SY2601 in EY liquid and rice solid media, resulting in the isolation and structure determination of 28 metabolites, including five new compounds, asperindopiperazines A-C (1-3), 5-methoxy-8,9-dihydroxy-8,9-deoxyaspyrone (21), and 12S-aspertetranone D (26). Structures of the new compounds were elucidated based on extensive NMR spectral analyses, HRESIMS data, optical rotation, ECD, and 13C NMR calculations. The new compound 12S-aspertetranone D (26) exhibited antibacterial activity against both methicillin-resistant Staphylococcus aureus and Escherichia coli with MIC values of 3.75 and 5 µg/mL, respectively.

Influence of Borassus flabellifer Endocarps Hydrolysate on Fungal Biomass and Fatty Acids Production by the Marine Fungus Aspergillus sp.

Polyunsaturated Fatty Acids (PUFAs) are important nutrients for human health. We aimed to evaluate the efficiency of marine water fungus Aspergillus sp. (Accession no: MZ505709) for lipid biosynthesis. The Yeast Extract Glucose (YEG) medium was supplemented with different concentration of Borassus flabellifer Endocarps Hydrolysate (BFEH; 1-5%) to evaluate the fungal biomass and its lipid accumulation. The combination of glucose and BFEH as carbon source increased the fresh weight (25.43 ± 0.33 g/L), dry weight (21.39 ± 0.77 g/L) and lipid yield (3.14 ± 0.09 g/L) of fungal biomass. The lipid content of dried fungal biomass has shown 91.08 ± 5.07 mg cod liver oil equivalents/g and 125.98 ± 5.96 mg groundnut oil equivalents/g biomass. GC-MS and NMR spectrometry analysis revealed the compounds involved in fatty acid metabolism and lipid signaling pathways along with the presence of linolenic acid. Interestingly, fungus grown in BFEH enriched medium has recorded the maximum amount of lipids with major fatty acid derivatives. Increase in the growth rate of Artemia franciscana was observed, when the extracted fungal lipid was supplemented as a food supplement. Therefore, this study suggests that marine fungal lipid may serve as potential natural compound as nutraceuticals and aquafeeds.

A New Brominated Isocoumarin from the Marine Starfish-Associated Fungus Aspergillus sp. WXF1904.

Based on the one strain many compounds strategy, a new brominated isocoumarin, 5-bromo-6,8-dihydroxy-3,7-dimethylisocoumarin (1), along with four new natural products, methyl 3-bromo-2,4-dihydroxy-6-methylbenzoate (2), methyl 2-bromo-4,6-dihydroxybenzoate (3), (E)-3-(3-bromo-4-hydroxyphenyl) acrylic acid (4) and 4-hydroxy-3-methyl-6-phenyl-2H-pyran-2-one (5), and four known compounds, methyl orsellinate (6), 4-hydroxy-3-methyl-6-(1-methyl-1-propenyl)-2H-pyran-2-one (7), pilobolusate (8) and cis-ferulic acid (9), were isolated from the ethyl acetate extract of the fungus Aspergillus sp. WXF1904 under the condition of adding bromine salt to the production medium. The structures of the new compounds were established by analysis of NMR and MS data. Compounds (1-9) were evaluated for inhibitory activity of acetylcholinesterase and pancreatic lipase, the new compound 1, known compounds 6 and 7 displayed weak inhibitory activity against acetylcholinesterase, compounds 2, 5, 7 and 8 showed weak inhibitory activity against pancreatic lipase.