Aspergillus Versicolor Research Articles

Chromones from the Tobacco Derived Fungus Aspergillus versicolor and Their Antiviral Activity

Chromones from the Tobacco Derived Fungus Aspergillus versicolor and Their Antiviral Activity

Bioactive Metabolite from Endophytic Aspergillus versicolor SB5 with Anti-Acetylcholinesterase, Anti-Inflammatory and Antioxidant Activities: In Vitro and In Silico Studies.

Endophytic fungi are a highly unpredictable group of microorganisms that can create a diverse range of secondary metabolites with biological activity. These metabolites enhance the host's ability to tolerate stress caused by various factors, such as disease, insects, pathogens, and herbivores. The secondary metabolites produced by endophytic fungi may have potential applications in agriculture, pharmacy, and medicine. The purpose of this study was to examine the anti-acetylcholinesterase activity of secondary metabolites extracted from endophytic fungi. Aspergillus versicolor SB5 was one of the many endophytic fungi isolated from Juncus rigidus and identified genetically with accession number ON872302. Our study utilized fermentation and microbial cultivation techniques to obtain secondary metabolites. During the course of our investigation, we isolated a compound called Physcion (C1) from the endophytic fungus Aspergillus versicolor SB5. We subsequently identified that C1 possesses inhibitory activity against COX-2 and LOX-1, with IC50 values of 43.10 and 17.54 µg/mL, respectively, making it an effective anti-inflammatory agent. Moreover, we found that C1 also exhibited potent anticholinesterase activity (86.9 ± 1.21%). In addition to these promising therapeutic properties, our experiments demonstrated that C1 possesses strong antioxidant capacity, as evidenced by its ability to scavenge DPPH, ABTS, O2 radicals, and NO and inhibit lipid peroxidation. To further investigate the molecular mechanisms underlying C1 pharmacological properties, we employed SwissADME web tools to predict the compound's ADME-related physicochemical properties and used Molecular Operating Environment and PyMOL for molecular docking studies.

Secondary metabolites from the Cordyceps-colonizing fungus Aspergillus versicolor ZJUTE2

Two new polyketides versicolorones A-B (1–2), one new diketopiperazine derivative aspergiamide B methyl ester (3), along with twenty known compounds 4–23, were obtained from the EtOAc extract of the Cordyceps-colonizing fungus Aspergillus versicolor ZJUTE2. The structures of 1–3 were established by detailed interpretation of the spectroscopic data and their absolute configurations were established by comparative analyses of the calculated and experimental ECD spectra. In the in-vitro bioassay, compounds 8 and 21 exhibited significant inhibitory activity against the Escherichia coli β-glucuronidase (EcGUS) with IC50 values of 54.73 ± 2.69 and 56.59 ± 1.77 μM, respectively.

Structure Revisions of Phenolic Bisabolane Sesquiterpenes and a Ferroptosis Inhibitor from the Marine-Derived Fungus Aspergillus versicolor YPH93.

Seven new phenolic bisabolane sesquiterpenoids (1-7), along with 10 biogenetically related analogues (8-17), were obtained from the deep-sea-derived fungus Aspergillus versicolor YPH93. The structures were elucidated based on extensive analyses of the spectroscopic data. Compounds 1-3 are the first examples of phenolic bisabolanes that contain two hydroxy groups attached to the pyran ring. The structures of sydowic acid derivatives (1-6 and 8-10) were carefully studied, leading to the structure revisions of six known analogues, including a revision of the absolute configuration for sydowic acid (10). All metabolites were evaluated for their effects on ferroptosis. Compound 7 exerted inhibition on erastin/RSL3-induced ferroptosis with EC50 values ranging from 2 to 4 μM, while it exhibited no effects on TNFα-induced necroptosis or H2O2-induced cell necrosis.

In Search of Clinical Markers: Indicators of Exposure in Dampness and Mold Hypersensitivity Syndrome (DMHS)

Potential markers were sought to diagnose mold hypersensitivity. Indoor air condensed water and human macrophage THP-1 test were applied to evaluate the buildings. Basophil activation tests (BAT) were conducted and mold-specific immunoglobulins (IgE, IgG, IgA, and IgD) were measured in study subjects' serum and feces. Exposed subjects reported markedly more symptoms from occupational air than controls. Basophils from exposed subjects died/lost activity at 225 times lower concentrations of toxic extracts from the target building than recommended in the common BAT protocol. Fecal IgG and IgD levels against Acrostalagmus luteoalbus and Aspergillus versicolor produced receiver operating curves (ROC) of 0.928 and 0.916, respectively, when plotted against the inflammation marker MRP8/14. Assaying serum immunoglobulin concentrations against the toxic Chaetomium globosum (MTAV35) from another building, a test control, did not differentiate study individuals. However, if liver metabolism produced the same core molecule from other Chaetomium globosum strains, this would explain the increased response in fecal immunoglobulins in the exposed. The altered immunoglobulin values in the samples of exposed when compared to controls revealed the route of mold exposure. The toxicity of indoor air condensed water samples, BAT and serology confirmed the severity of symptoms in the target building's employees, supporting earlier findings of toxicity in this building.

Effect of Culture Conditions of Lophocereus marginatus Endophytic Fungi on Yield and Anticancer and Antioxidant Activities

Culture conditions affect the production of secondary metabolites in endophytic fungi. Therefore, the aim of the present study was to evaluate the yield and anticancer and antioxidant activity of endophytic fungi extracts from the cactus Lophocereus marginatus, under different culture conditions. The strains Penicillium citrinum, Aspergillus versicolor, Metarhizium anisopliae, and Cladosporium sp. were fermented in different culture media (potato dextrose agar, Czapeck broth, and malt broth), types of inoculums (spore or mycelium), and shaking conditions (150 rpm or static) for one week. Methanol extracts were obtained from mycelia, which was followed by determining their yields and evaluating their effect on L5178Y-R murine lymphoma cells growth and human peripheral blood mononuclear cells (PBMCs) viability, using the 3-[4,5dimethylthiazol-2-yl]2,5-diphenyl tetrazolium bromide reduction colorimetric assay. In addition, antioxidant activity was determined by the 2,2-diphenyl-1-picrylhydrazyl test. We determined the half-maximal inhibitory concentration (IC50) values of tumor cell growth inhibition, the selectivity index (SI), and the antioxidant activity, as compared with the healthy cells control. The best yields were obtained with the Czapeck broth medium in all the evaluated strains, reaching values of 50.3%. Of the 48 extracts evaluated, only seven significantly (p < 0.01) inhibited tumor cell growth (IC50 < 250 µg/mL). A. versicolor extract showed the highest anticancer activity, after culturing spores (IC50 = 49.62 µg/mL; SI = 15.8) or mycelium (IC50 = 69.67 µg/mL; SI = 12.2) in malt broth, under static conditions. Extracts did not present significant antioxidant activity. In conclusion, we showed that culture conditions influenced the anticancer activity of L. marginatus endophytic fungi.

Oscillatoria limnetica Mediated Green Synthesis of Iron Oxide (Fe2O3) Nanoparticles and Their Diverse In Vitro Bioactivities.

Iron oxide nanoparticles (Fe2O3-NPs) were synthesized using Oscillatoria limnetica extract as strong reducing and capping agents. The synthesized iron oxide nanoparticles IONPs were characterized by UV-visible spectroscopy, Fourier transform infrared (FTIR), X-ray diffractive analysis (XRD), scanning electron microscope (SEM), and Energy dispersive X-ray spectroscopy (EDX). IONPs synthesis was confirmed by UV-visible spectroscopy by observing the peak at 471 nm. Furthermore, different in vitro biological assays, which showed important therapeutic potentials, were performed. Antimicrobial assay of biosynthesized IONPs was performed against four different Gram-positive and Gram-negative bacterial strains. E. coli was found to be the least suspected strain (MIC: 35 µg/mL), and B. subtilis was found to be the most suspected strain (MIC: 14 µg/mL). The maximum antifungal assay was observed for Aspergillus versicolor (MIC: 27 µg mL). The cytotoxic assay of IONPs was also studied using a brine shrimp cytotoxicity assay, and LD50 value was reported as 47 µg/mL. In toxicological evaluation, IONPs was found to be biologically compatible to human RBCs (IC50: >200 µg/mL). The antioxidant assay, DPPH 2,2-diphenyl-1-picrylhydrazyly was recorded at 73% for IONPs. In conclusion, IONPs revealed great biological potential and can be further recommended for in vitro and in vivo therapeutic purposes.

Isolation of the Main Pathogens Causing Postharvest Disease in Fresh Angelica sinensis during Different Storage Stages and Impacts of Ozone Treatment on Disease Development and Mycotoxin Production.

Angelica sinensis, a Chinese herbal medicine, is susceptible to molds during storage, reducing its quality, and even generating mycotoxins with toxic effects on human health. Fresh A. sinensis was harvested from Min County of Gansu Province in China and kept at room temperature. Naturally occurring symptoms were observed during different storage stages. Molds were isolated and identified from the diseased A. sinensis using morphological and molecular biology methods. The impact of ozone treatment on postharvest disease development and mycotoxin production was investigated. The results indicated that A. sinensis decay began on day 7 of storage and progressed thereafter. Nine mold species were isolated and characterized: day 7, two Mucormycetes; day 14, Clonostachys rosea; day 21, two Penicillium species and Aspergillus versicolor; day 28, Alternaria alternata and Trichoderma atroviride; and day 49, Fusarium solani. Ozone treatment markedly inhibited the development of postharvest disease and the mycotoxin production (such as, patulin, 15-acetyl-deoxynivalenol, and sterigmatocystin) in the rotten tissue of A. sinensis inoculated with the nine isolates.

Deep Paleoproteotyping and Microtomography Revealed No Heart Defect nor Traces of Embalming in the Cardiac Relics of Blessed Pauline Jaricot.

Scientific examination of the heart of Blessed Pauline Jaricot-a French missionary figure-was carried out in 2022. As tandem mass spectrometry proteotyping has proven to be valuable to obtain the broad taxonomic repertoire of a given sample without any a priori information, we aimed at exploring the conditions of preservation of the relics and possible conditions of death. Metaproteomics and high-resolution microtomography imaging approaches were combined. A dataset comprising 6731 high-resolution MS/MS spectra was acquired and 968 of these spectra could be assigned to specific peptidic biomolecules. Based on the taxonomical information encompassed by the identified peptide sequences, 5 phyla were identified amongst eukaryota (94% of the biomass): Ascomycota (55%), with the species Aspergillus versicolor, Trichophyton mentagrophytes and Aspergillus glaucus, corresponding to expected cadaverous fungal flora; Chordata (42%), represented by a unique species, Homo sapiens; Streptophyta (3%); and Arthropoda (traces). Bacteria (6% of the biomass) were poorly represented. No trace of embalming substance could be retrieved, nor any pathogens. Imaging evidenced no heart defect nor embalming traces. No evidence that was inconsistent with natural and spontaneous conservation could be retrieved. This study prefigures the power of modern molecular techniques such as paleoproteotyping coupled to microtomography to gain insight into historical relics.

Mechanisms of airway disease development following inhalation exposure to indoor fungal contaminants Stachybotrys chartarum and Aspergillus versicolor

Fungi are found ubiquitously in the environment and chronic repeated exposures have been associated with pulmonary and cognitive effects in humans. Previous studies have shown that two prominent indoor fungal contaminants, Stachybotrys chartarum and Aspergillus versicolor, are associated with allergic airway disease, airway remodeling, and Th2-mediated immunological effects. To understand the mechanisms of disease development, genetically modified murine models are an invaluable tool.

Kojicone C, new anti-inflammatory polyketide from insect-associated fungus Aspergillus versicolor

The chemical study of EtOAc extracts of the insect-associated fungus Aspergillus versicolor led to the isolation of a new polyketide kojicone C ( 1 ) and its precursor (2 R ,4 R )−4-hydroxy-5-methoxy-2,4-dimethyl-2-[(2 R )−2-methylbutyryloxy]−5-cyclohexen-1,3-dione ( 2 ). Kojicone C belongs to the rare tricycle cyclohexen-1,3-dione adduct class. The structures of 1 and 2 were determined by HRESIMS (high resolution electrospray ionization mass spectroscopy) data, 1D and 2 D NMR (nuclear magnetic resonance) spectroscopic analysis, ECD (electronic circular dichroism) calculations, and CD exciton chirality method. Compound 1 exhibited moderate anti-inflammatory activities that reduced LPS-induced NO production of RAW 264.7 cells (IC 50 = 35.8 μ Μ). It could also significantly inhibit LPS-induced mRNA expression of TNF- α , IL-1 β , and IL-6 of this cell line at a concentration of 20 μ Μ. • One new polyketide was isolated from Aspergillus versicolor . • The new compound is a rare tricycle cyclohexen-1,3-dione adduct. • The new compound exhibited potent anti-inflammatory activities.

Bioactivity and Metabolome Mining of Deep-Sea Sediment-Derived Microorganisms Reveal New Hybrid PKS-NRPS Macrolactone from Aspergillus versicolor PS108-62

Despite low temperatures, poor nutrient levels and high pressure, microorganisms thrive in deep-sea environments of polar regions. The adaptability to such extreme environments renders deep-sea microorganisms an encouraging source of novel, bioactive secondary metabolites. In this study, we isolated 77 microorganisms collected by a remotely operated vehicle from the seafloor in the Fram Strait, Arctic Ocean (depth of 2454 m). Thirty-two bacteria and six fungal strains that represented the phylogenetic diversity of the isolates were cultured using an One-Strain-Many-Compounds (OSMAC) approach. The crude EtOAc extracts were tested for antimicrobial and anticancer activities. While antibacterial activity against methicillin-resistant Staphylococcus aureus (MRSA) and Enterococcus faecium was common for many isolates, only two bacteria displayed anticancer activity, and two fungi inhibited the pathogenic yeast Candida albicans. Due to bioactivity against C. albicans and rich chemical diversity based on molecular network-based untargeted metabolomics, Aspergillus versicolor PS108-62 was selected for an in-depth chemical investigation. A chemical work-up of the SPE-fractions of its dichloromethane subextract led to the isolation of a new PKS-NRPS hybrid macrolactone, versicolide A (1), a new quinazoline (-)-isoversicomide A (3), as well as three known compounds, burnettramic acid A (2), cyclopenol (4) and cyclopenin (5). Their structures were elucidated by a combination of HRMS, NMR, [α]D, FT-IR spectroscopy and computational approaches. Due to the low amounts obtained, only compounds 2 and 4 could be tested for bioactivity, with 2 inhibiting the growth of C. albicans (IC50 7.2 µg/mL). These findings highlight, on the one hand, the vast potential of the genus Aspergillus to produce novel chemistry, particularly from underexplored ecological niches such as the Arctic deep sea, and on the other, the importance of untargeted metabolomics for selection of marine extracts for downstream chemical investigations.

Targeted isolation of antitubercular cycloheptapeptides and an unusual pyrroloindoline-containing new analog, asperpyrroindotide A, using LC–MS/MS-based molecular networking

Further insights on the secondary metabolites of a soft coral-derived fungus Aspergillus versicolor under the guidance of MS/MS-based molecular networking led to the isolation of seven known cycloheptapeptides, namely, asperversiamides A–C (1–3) and asperheptatides A–D (4–7) and an unusual pyrroloindoline-containing new cycloheptapeptide, asperpyrroindotide A (8). The structure of 8 was elucidated by comprehensive spectroscopic data analysis, and its absolute configuration was determined by advanced Marfey’s method. The semisynthetic transformation of 1 into 8 was successfully achieved and the reaction conditions were optimized. Additionally, a series of new derivatives (10−19) of asperversiamide A (1) was semi-synthesized and their anti-tubercular activities were evaluated against Mycobacterium tuberculosis H37Ra. The preliminary structure−activity relationships revealed that the serine hydroxy groups and the tryptophan residue are important to the activity.

TUBB1, TUBA4A and MAPK as Indicators of Die-Back Degenerative Central Nervous System Disease in Patients Sickened by Specific Exposure to the Interior Environment of Water- Damaged Buildings

Beginning with Possible Estuarine Associated Syndrome (PEAS) in 1998 and followed by numerous studies on similar types of exposures, patients with illness associated with biotoxin exposure routinely have had symptom rosters dominated by executive cognitive dysfunction, including recent memory deficits, difficulty in concentration, difficulty with word finding, decreased assimilation of new knowledge, confusion and disorientation, tremors, headaches, vertigo, and unusual pains. In addition, tremors, headaches, vertigo, unusual pains and metallic taste. Toxins involved variously have included those found in toxin-forming dinoflagellates, including Pfiesteria and ciguatera; cyanobacteria, including Microcystis, Cylindrospermopsis and Lyngbya wollei; post Lyme syndrome and Babesia; as well as organisms found in damp buildings, including Aspergillus versicolor and A. penicillioides, Stachybotrys chartarum, Chaetomium globosum, Wallemia sebi, Actinobacteria species, especially Corynebacteria tuberculostearicum and Propionibacterium acnes; as well as bacterial endotoxins and beta-glucans. These illnesses have been called chronic inflammatory response syndromes (CIRS). The mechanism of neurologic findings has remained elusive despite studies showing successful treatment with intranasal vasoactive intestinal polypeptide (VIP). Neurocognitive testing has only been performed in PEAS patients, showing profound deficits in learning and higher cognitive functioning. These CIRS patients have had brain imaging without consistent findings, including MRI, EEG, and CT of the brain. NeuroQuant has shown findings that fit a “fingerprint” found in patients with specific causation and confirmed exposure to Actinobacteria and endotoxins. Fungal exposure shows disproportionate enlargement due to interstitial edema in the forebrain parenchyma and cortical grey, with a diminished caudate nucleus size. These findings have not been found in controls. The recent inclusion of transcriptomic studies using GENIE has confirmed that specific causation can be identified for Actinobacteria (48% of total confirmed cases), bacterial endotoxins (28%) and fungi (7%) in CIRS patients. The combination of NeuroQuant and GENIE has implicated excessive production of cytoskeletal tubulin genes TUBA4A and TUBB1 as risk factors for specific fingerprints for die-back degenerative central nervous system (CNS) injury in patients with illnesses associated variously with exposure to Actinobacteria, fungi and endotoxins. This study seeks to implicate a causal abnormality of excessive expression of tubulin genes TUBA4A and TUBB1. Given the role of these genes in die-back CNS degenerative diseases, such as Alzheimer’s, amyotrophic lateral sclerosis and Parkinson’s disease, and anecdotal successful treatment of CIRS patients with elevated TUBA4A and TUBB1, we suggest the possibility of treatment of tubulin excess may have a role in clinical improvement seen in die-back CNS degenerative diseases. Elevated levels of MAPK are also risk factors when combined with elevated levels of TUBA4A and TUBBI.

Antimicrobial pontential of fungal endophytes obtained from &lt;em&gt;Avicennia Marina&lt;/em&gt; and &lt;em&gt;Excoecaria Agallocha&lt;/em&gt; in Negombo

Mangroves are halophytic plants that are typically found along the intra-tidal regions. The coastal ecosystem of mangroves in Sri Lanka hold one of the most unique mangrove species that are yet to be studied. These ecosystems provide shelter to various organisms including humans. Fungal endophytes present in these plants have many medicinal properties that can be utilized in the development of drugs. Antibiotic resistance is a major concern in the pharmaceutical industry, hence alternative plant-based drugs with fewer costs and side effects show great potential. In the current investigation, the main objective was to test for antibacterial properties of fungal endophytes isolates from two mangroves: Avicennia marina and Excoecaria agallocha against Staphylococcus aureus (ATCC 25923) and Escherichia coli (ATCC 25922). In both mangrove species, three endophytic species were identified based on their morphological characteristics: Aspergillus niger, Penicillium viticola and Aspergillus versicolor. Only P. viticola was obtained from E. agallocha. Based on the results obtained from antibiotic susceptibility tests, A. niger (stem) was the most active and showed the best bacteriostatic and bactericidal activity against both S. aureus and E. coli. P. viticola produced viable results (mean zone of inhibition of 21 mm against S. aureus) but were not as productive as A. niger. This study indicated that these endophytic fungi held medicinal properties that can be further evaluated.

Anti-Tobacco Mosaic Virus Indole Alkaloids from the Nicotiana tabacum-Derived Fungus Aspergillus versicolor

To explore the active metabolites from Aspergillus, we have discovered five undescribed tricyclic or tetracyclic indole alkaloids aspergillines K–O (1–5) and four known analogues (6–9) from the strain of Nicotiana tabacum-derived Aspergillus versicolor YNCA0363. The α-amino group derived from tryptophan in isolates 1–6 was changed to an oxygen atom, which is uncommon in the reported analogues. To the best of our knowledge, 4 and 5 represent the first examples of indole alkaloids possessing an isochromene moiety. Compounds 1 and 3 showed obvious anti-tobacco mosaic virus (TMV) activities with inhibition rates of 43.6% ± 2.2 and 58.5% ± 2.8 at the concentration of 20 μM, respectively, which were significantly higher than that of the positive control. Compound 3 also showed excellent potential of inducing systemic acquired resistance. Furthermore, compound 3 can increase the activities of two reactive oxygen species-scavenging enzymes, peroxidase and superoxide dismutase, and enhance tobacco resistance to TMV. Molecular docking results showed that compound 3 exhibits a stronger interaction with TMV-helicase, and showed higher antiviral activities than others. This work revealed that the above indole alkaloids, especially 3, could induce resistance in tobacco plants against the viral pathogen TMV.

Structural characterization and immunomodulatory activity of an exopolysaccharide from marine-derived Aspergillus versicolor SCAU141

Until now, relatively little is known about marine-derived fungal polysaccharides and their activities. Exopolysaccharide AVP141-A was isolated from the broth of marine-derived fungus Aspergillus versicolor SCAU141 and purified by Diethylaminoethyl-Sepharose Fast Flow and Sephadex G-100. The structural characteristics of AVP141-A was studied by chemical analysis together with high-performance gel permeation chromatography, ion chromatography, Fourier-transform infrared spectroscopy, gas chromatography–mass spectrometry and nuclear magnetic resonance spectroscopy. The results showed that AVP141-A with the molecular weight of 5.10 kDa was mainly composed of →4)-α-D-Glcp-(1→, branched by α-D-Glcp-(1→ and →6)-α-D-Glcp-(1→ at C-6 positions of the glucan backbone. In particular, sulfate ester (approximately 3.62 %) was found in AVP141-A, which was frequently considered to occur in marine-derived microbial polysaccharides rather than other microbial polysaccharides. Furthermore, AVP141-A significantly enhanced the activity of the inflammatory factors NO, COX-2 and TNF-α in RAW264.7 macrophages by activating the MAPK/p38 and NF-κB/p65 pathways. In addition, metabolomic analysis revealed that most of the pathways with significant changes in RAW264.7 macrophages treated with AVP141-A were amino acid-related pathways, and arginine was the characteristic metabolite. In conclusion, this study identified AVP141-A as a marine fungus-derived sulfated exopolysaccharide with potential for development as an immune activator.

New application of Aspergillus versicolor in promoting plant growth after suppressing sterigmatocystin production via genome mining and engineering.

Aspergillus genus is a key component in fermentation and food processing. However, sterigmatocystin (STE)-a mycotoxin produced by several species of Aspergillus-limits the use of some Aspergillus species (such as Aspergillus versicolor, Aspergillus inflatus, and Aspergillus parasiticus) because of its toxicity and carcinogenicity. Here, we engineered an STE-free Aspergillus versicolor strain based on genome mining techniques. We sequenced and assembled the Aspergillus versicolor D5 genome (34.52 Mb), in which we identified 16 scaffolds and 54 biosynthetic gene clusters (BGCs). We silenced cytochrome P450 coding genes STC17 and STC27 by insertional inactivation. The production of STE in the Δstc17 mutant strain was increased by 282% but no STE was detected in the Δstc27 mutant. Metabolites of Δstc27 mutant exhibited growth-promoting effect on plants. Our study makes significant progress in improving the application of some Aspergillus strains by restricting their production of toxic and carcinogenic compounds.

Atmospheric Corrosion of Copper by an Acid-Producing Aspergillus versicolor in the Presence of Chloride

Atmospheric Corrosion of Copper by an Acid-Producing Aspergillus versicolor in the Presence of Chloride

Metagenomics of Toenail Onychomycosis in Three Victorian Regions of Australia.

Onychomycosis is a fungal disease of the nail that is found worldwide and is difficult to diagnose accurately. This study used metagenomics to investigate the microbiology of 18 clinically diagnosed mycotic nails and two normal nails for fungi and bacteria using the ITS2 and 16S loci. Four mycotic nails were from Bass Coast, six from Melbourne Metropolitan and eight from Shepparton, Victoria, Australia. The mycotic nails were photographed and metagenomically analysed. The ITS2 sequences for T. rubrum and T. interdigitale/mentagrophytes averaged over 90% of hits in 14/18 nails. The high abundance of sequences of a single dermatophyte, compared to all other fungi in a single nail, made it the most likely infecting agents (MLIA). Trichophyton rubrum and T. interdigitale/mentagrophytes were found in Bass Coast and Shepparton while only T. interdigitale/mentagrophytes was found in Melbourne. Two nails with T. interdigitale/mentagrophytes mixed with high abundance non-dermatophyte moulds (NDMs) (Aspergillus versicolor, Acremonium sclerotigenum) were also observed. The two control nails contained chiefly Fusarium oxysporum and Malassezia slooffiae. For bacteria, Staphylococcus epidermidis was in every nail and was the most abundant, including the control nails, with an overall mean rate of 66.01%. Rothia koreensis, Corynebacterium tuberculostearicum, and Brevibacterium sediminis also featured.