Detection of Ophidiomyces and Nannizziopsis spp. in the dermal lesions of free-roaming native snakes in Taiwan.

Chrysosporium-related fungi refers to an assemblage of fungi belonging to the Nannizziopsis, Ophidiomyces, and Paranannizziopsis genera. Chrysosporium-related fungi infection results in various skin lesions, such as necrosis and ulcers, in both captive and free-roaming reptiles. To update the prevalence of ophidiomycosis in Taiwan, which was first detected in 2019, we conducted a large-scale ecological survey of free-roaming native snakes with skin lesions in Taiwan. To the best of our knowledge, this is the first study on Chrysosporium-related fungi prevalence in Southeast Asia. Fungal samples collected from the skin lesions of snakes were cultured and subjected to morphological, histopathological, and molecular analyses. We examined 2382 free-roaming snakes representing 42 snake species; among them, 132 (5.54%) had skin lesions. Ten (0.42%) snakes, representing four species, tested positive for Ophidiomyces (five snakes, four species) or a possibly novel Nannizziopsis species (five snakes, three species). The infected snakes were generally healthy, with mild clinical signs. The low prevalence rate, mild clinical signs, and broad pathogen/host range suggest that Ophidiomyces and Nannizziopsis are endemic to Southeast Asia. The newly reported presence of Nannizziopsis in free-roaming snakes suggests the need for modifying the currently used surveillance strategy.

We detected species of Ophidiomyces and Nannizziopsis in free-roaming snakes in Taiwan. The low prevalence and mild clinical signs suggest they may be endemic in Taiwan. The finding of Nanninzziopsis in free-roaming snakes indicates potential cross-host infection of onygenalean fungus in reptiles.

Chrysosporium articulatum mimicking Trichophyton spp. infection in a cat: a case presentation and literature review.

BACKGROUND: Dermatophytosis is a common skin infection of cats and many other animals. A reliable diagnosis is crucial because of the zoonotic potential of dermatophytes. The routine mycological diagnostic procedures for dermatophytosis are widely known, but in the case of some isolates, identification based on phenotypic characteristics may be incorrect. Infections caused by Chrysosporium spp. are usually described in reptiles, but in other animals they are uncommon. CASE PRESENTATION: This study presents a description of a cat with dermatological lesions, that was mistakenly diagnosed with Trichophyton spp. dermatophytosis. Clinical material for mycological examination was collected from alopecic areas on the back of the neck, the ventral abdomen, and the hindlimbs. The initial identification based on phenotypic properties indicated Trichophyton spp. The result of the MALDI-ToF MS allowed the exclusion of the Trichophyton genus. Ultimately, the correct identification as Chrysosporium articulatum was obtained based on the sequencing of ribosomal genes. CONCLUSIONS: Interpretation of the results of the mycological examination of samples collected from animals' skin or hair shafts is always challenging. Thus, careful consideration of the primary cause of the clinical lesions observed on the skin is mandatory, and the culture results are worth supporting by molecular methods.

Chrysosporium: A rare cause of allergic fungal rhinosinusitis.

Allergic fungal rhinosinusitis (AFRS) forms a significant group of patients presenting with the commonest health problem encountered in rhinology. Patients commonly present with typical symptoms of sinusitis, and the diagnosis is often made after imaging and/or intraoperatively. Infections caused by Chrysosporium species are very rare and are very rarely been reported to cause sinusitis in humans. Usually, human chrysosporial infections are mild and unmarked by symptoms. We report a rare case of allergic fungal sinusitis (AFS) caused by Chrysosporium species in a 41-year-old male with the history of diabetes mellitus.

Use of Chrysosporium/carbon nanotubes for preconcentration of ultra-trace cadmium levels from various samples after extensive studies on its adsorption properties.

Carbon nanotubes were used to immobilize Chrysosporium fungus for building an adequate adsorbent to be used as an desirable sorbent for preconcentration and measurement of cadmium ultra-trace levels in various samples. After characterization, the potential of Chrysosporium/carbon nanotubes for the sorption of Cd(II) ions was scrutinized by the aid of central composite design, and comprehensive studies of sorption equilibrium, kinetics and thermodynamic aspects were accomplished. Then, the composite was utilized for preconcentration of ultra-trace cadmium levels, by a mini-column packed with Chrysosporium/carbon nanotubes, before its determination with ICP-OES. The outcomes vouchsafed that (i) Chrysosporium/carbon nanotube has a high tendency for selective and rapid sorption of cadmium ion, at pH 6.1, and (ii) kinetic, equilibrium, and thermodynamic studies showed a high affinity of the Chrysosporium/carbon nanotubes for cadmium ion. Also, the outcomes displayed that cadmium can quantitatively be sorbed at a flow speed lesser than 7.0 mL/min and a 1.0 M HCl solution (3.0 mL) was sufficient to desorbe the analyte. Eventually, preconcentration and measurement of Cd(II) in different foods and waters were successfully accomplished with good accuracy, high precision (RSDs ≤5.65%), and low limit of detection (0.015 µg/L).

Nannizziopsis guarroi has prolonged environmental persistence on clinically relevant substrates.

OBJECTIVE: To determine the environmental persistence of Nannizziopsis guarroi on clinically relevant solid and aqueous substrates. SAMPLE: 2 molecularly confirmed isolates of N guarroi obtained from clinical cases of dermatomycosis in bearded dragons (Pogona vitticeps). PROCEDURES: 3 concentrations (1 McFarland, 1:10 McFarland, and 1:100 McFarland) of fungal suspension were exposed to 7 sterilized solid substrates (fabric aquarium liner, wood mulch, sand, hard plastic, glass, cotton, and stainless steel) and 2 sterilized aqueous substrates (distilled water, saline solution [0.9% NaCl]). Biological replicates were performed for the contamination of the solid substrates. On days 1, 3, and 14 after contamination, the substrates were sampled for fungal culture with technical repeat. Fungal cultures were incubated at room temperature for 10 days and then evaluated for fungal growth. RESULTS: Data from wood mulch were not evaluated because of plate contamination. Overall, the ability to culture N guarroi from solid substrates was isolate, time, and fungal concentration dependent. Viable fungus was isolated from fabric aquarium liner and glass on day 1 and days 1 and 3, respectively. N guarroi was cultured from all other solid substrates at day 14 from at least 1 isolate and/or fungal concentration. Viable N guarroi was isolated from both aqueous substrates at day 14, regardless of isolate or fungal concentration. CLINICAL RELEVANCE: The environmental persistence of N guarroi should be considered when treating lizards infected with this fungus. Fomites may contribute to the contagious nature of this pathogen and environmental disinfection should be performed to reduce transmission.

Cyclopiazonic Acid and Okaramine Analogues, Including Chlorinated Compounds, from Chrysosporium undulatum YT-1.

Eight new cyclopiazonic acid (1-8) and five new okaramine (9-13) alkaloids together with 13 known compounds were isolated from the fungus Chrysosporium undulatum YT-1. Compounds 2, 4, 5, 7, 10, 11, and 13 were chlorinated indole alkaloids. The structures of compounds 1-13 were elucidated by HRESIMS and NMR spectroscopic data. Their relative and absolute configurations were established by J-based configuration analysis, NOESY, NOEDIFF experiments, ECD spectroscopic data, and biogenetic considerations. Compound 4 inhibited the growth of Bacillus subtilis with an MIC value of 6.3 µg/mL. Compounds 9-11 exhibited strong insecticidal capacity against the third instar larvae of silkworm and cotton bollworm (LD50: ≤7.56 µg/g). At 40 µM, compound 1 showed obvious neuroprotection to the PC12 cells with 6-OHDA treatment.

Neochrysosporazines: Precursor-Directed Biosynthesis Defines a Marine-Derived Fungal Natural Product P-Glycoprotein Inhibitory Pharmacophore.

Upregulation of ATP binding cassette (ABC) transporter efflux pumps (i.e. P-glycoprotein, P-gp) can impart multidrug resistance, rendering many chemotherapeutics ineffective and seriously limiting treatment regimes. While ABC transporters remain an attractive target for therapeutic intervention, the development of clinically useful small-molecule inhibitors has proved challenging. In this report, we describe the structure-activity relationship (SAR) analysis of a newly discovered P-gp inhibitory pharmacophore, phenylpropanoid piperazine chrysosporazines, produced by co-isolated marine-derived fungi. In the absence of any total syntheses, we apply an innovative precursor-directed biosynthesis strategy that successfully repurposed fungal biosynthetic output, allowing us to isolate, characterize, and identify the structurally diverse neochrysosporazines A-Q. SAR analysis utilizing all known (and new) neochrysosporazines, chrysosporazines, and azachrysosporazines, plus semi-synthetic analogues, established the key structure requirements for the P-gp inhibitory pharmacophore, and, in addition, identified non-essential sites that allow for the design of affinity and other conjugated probes.

Koch's postulates: Confirming Nannizziopsis guarroi as the cause of yellow fungal disease in Pogona vitticeps.

Nannizziopsis guarroi is an ascomycete fungus associated with a necrotizing dermatitis in captive green iguanas (Iguana iguana) and bearded dragons (Pogona vitticeps) across both Europe and North America. Clinical signs of the disease include swelling and lesion formation. Lesions develop from white raised bumps on the skin and progress into crusty, yellow, discolored scales, eventually becoming necrotic. The clinical signs are the basis of a colloquial name yellow fungal disease (YFD). However, until now, N. guarroi has not been confirmed as the primary agent of the disease in bearded dragons. In this experiment, we fulfill Koch's postulates criteria of disease, demonstrating N. guarroi as the primary agent of YFD in bearded dragons.

Precursor-Directed Biosynthesis Mediated Amplification of Minor Aza Phenylpropanoid Piperazines in an Australian Marine Fish-Gut-Derived Fungus, Chrysosporium sp. CMB-F214.

Chemical analysis of an M1 agar plate cultivation of a marine fish-gut-derived fungus, Chrysosporium sp. CMB-F214, revealed the known chrysosporazines A-D (11-14) in addition to a suite of very minor aza analogues 1-6. A microbioreactor (MATRIX) cultivation profiling analysis failed to deliver cultivation conditions that significantly improved the yields of 1-6; however, it did reveal that M2 agar cultivation produced the new natural product 15. A precursor-directed biosynthesis strategy adopting supplementation of a CMB-F214 M1 solid agar culture with sodium nicotinate enhanced production of otherwise inaccessible azachrysposorazines A1 (1), A2 (2), B1 (3), C1 (4), C2 (5) and D1 (6), in addition to four new chrysosporazines; chrysosporazines N-P (7-9) and spirochrysosporazine A (10). Structures inclusive of absolute configurations were assigned to 1-15 based on detailed spectroscopic and chemical analyses, and biosynthetic considerations. Non-cytotoxic to human carcinoma cells, azachrysosporazies 1-5 were capable of reversing doxorubicin resistance in P-glycoprotein (P-gp)-overexpressing human colon carcinoma cells (SW620 Ad300), with optimum activity exhibited by the C-2' substituted analogues 3-5.

Biosorption effect of Aspergillus niger and Penicillium chrysosporium for Cd- and Pb-contaminated soil and their physiological effects on Vicia faba L.

Phytoremediation is an important solution to soil pollution management. The goal of this study is to determine the biosorption ability of the two selected fungi (Aspergillus niger and Penicillium chrysosporium) under heavy metal stress on faba bean plants. The fungal strains produced phytohormones, siderophore, ACC deaminase, and secondary metabolites. The biosorption capacity of A. niger and P. chrysosporium was 0.09 and 0.06 mg g-1 and 0.5 and 0.4 mg g-1 in media containing Cd and Pb, respectively. Fourier transform infrared spectroscopy of the fungal cell wall show primary functional groups like hydroxyl, amide, carboxyl, phosphoryl, sulfhydryl, and nitro. Therefore, A. niger and P. chrysosporium were inoculated to soils, and then the faba bean seeds were sown. After 21 days of sowing, the plants were irrigated with water to severe as control, with 100 mg L-1 of Cd and 200 mg L-1 of Pb. The results show that Cd and Pb caused a significant reduction in morphological characteristics, auxin, gibberellins, photosynthetic pigments, minerals content, and antioxidant enzymes as compared to control plants but caused a substantial boost in abscisic acid, ethylene, electrolyte leakage, lipid peroxidation, glutathione, proline, superoxide dismutase, secondary metabolites, and antioxidant capacity. In inoculated plants, metal-induced oxidative stress was modulated by inhibiting the transport of metal and decreased electrolyte leakage and lipid peroxidation. Finally, the inoculation of endophytic fungi contributed actively to the absorption of heavy metals and decreased their content in soil and plants. This could be utilized as an excellent technique in the fields of heavy metal-contaminated sustainable agriculture.

Epidemiological aspects of superficial fungal infections in Koranic schools in two localities of Senegal (Thies and Touba).

BACKGROUND: In developing countries, superficial fungal infections (SFI) are endemic and cause a therapeutic problem because of the duration and cost of treatment. Community living and promiscuity are key factors in the direct or indirect transmission and spread of these diseases. OBJECTIVES: The objective was to study the epidemiological aspects of SFI, among koranic school children in two localities in Senegal. PATIENTS/METHODS: School koranic students were recruited in Thies and Touba. Diagnosis of fungal diseases was carried out using conventional techniques (microscopic examination and culture). RESULTS: Among 210 children, the overall prevalence of SFI was 25.71%, with 27.63% in Touba and 20.68% in Thiès. The clinical lesions were epidermophytosis (0.5%), intertrigo (0.9%), palmoplantar keratoderma (KPP) (0.9%), onychomycosis (7.7%) and tinea capitis (TC) (90%). The species responsible for the SFI were Trichophyton soudanense (85.18%), Microsporum audouinii langeronii (9.25%), Trichophyton rubrum (3.70%) and Chrysosporium keratinophilum (1.85%). The prevalence of infection was higher among boys (85.18%). CONCLUSION: Superficial fungal infections are prevalent in koranic school children and attention should be given to non-dermatophytic species that could be responsible for SFI.

Fir bark modified by Phanerodontia chrysosporium: A low-cost amendment for cd-contaminated water and agricultural soil.

In this study, a modified fir barks (MFB) was prepared by mixing fir barks (FB) and white-rot fungi (Phanerodontia chrysosporium) under aerobic fermentation. The potential of MFB for Cd2+ adsorption was investigated by batch experiments combined with kinetic, isotherm, and thermodynamics analyses. The results revealed that the modification greatly increased the porous structures on the surfaces of fir barks and the surface area of MFB was much higher than that of FB. As a result, the adsorption capacity of Cd2+ on MFB (17.4 mg g-1) was more than two times higher than that on FB (7.2 mg g-1), and the adsorption of Cd2+ on MFB was controlled by physisorption and chemisorption. The immobilization of Cd by MFB in a contaminated agricultural soil was also investigated. The effect of MFB on the bioavailability of Cd was investigated using a leaching test (the European standard EN 12457-2) combined with a typical sequential extraction procedure (the community bureau of reference, BCR). The experimental results showed that the Cd leachability was reduced by 71% when the added MFB dosage was 30 mg g-1. Besides, the MFB amendment could transform Cd from unstable geochemical fractions into more stable fractions. In total, the MFB, as a chemical-free and eco-friendly material, could be potentially employed for in-situ remediation of Cd-contaminated agricultural soils.

Adiaspore development and morphological characteristics in a mouse adiaspiromycosis model.

Lesions of adiaspiromycosis, a respiratory disease affecting wild animals, have been found mainly in dead mammals and free-living mammals captured for surveillance. No report has described an investigation of adiaspore formation progress in the lung. After establishing an experimental mouse model of intratracheal adiaspiromycosis infection with the causative agent Emmonsia crescens, we observed adiaspore development. The spores grew and reached a plateau of growth at 70 days post-infection. The median adiaspore diameter showed a plateau of around 40 µm. The characteristic three-layer cell-wall structure of adiaspores was observed in the lung at 70 days post-infection. We examined infection with a few spores, which revealed that adiaspores in the mouse lung progressed from intratracheal infection of at least 400 spores. Moreover, we developed adiaspores in vitro by culture in fetal bovine serum. Although most spores broke, some large spores were intact. They reached about 50 µm diameter. Thick cell walls and dense granules were found as common points between in vitro adiaspores and in vivo adiaspores. These models are expected to be useful for additional investigations of E. crescens adiaspores and adiaspiromycosis.

Intraspecific Diversity and Taxonomy of Emmonsia crescens.

Emmonsia crescens is known as an environmental pathogen causing adiaspiromycosis in small rodents. As the generic name Emmonsia is no longer available for this species, its taxonomic position is re-evaluated. The intraspecific variation of Emmonsia crescens was analyzed using molecular, morphological, and physiological data, and the relationship between frequency of adiaspiromycosis and body temperature of host animals was explored. A North American and a pan-global lineage could be discerned, each with subclusters at low genetic distance. European strains produced the classical type of very large adiaspores, while in the North American lineage adiaspores relatively small, resembling the broad-based budding cells of Blastomyces. Members of the closely related genus Emergomyces may exhibit large, broad-based in addition to small, narrow-based budding cells. We conclude that the morphology of the pathogenic phase in these fungi differs gradationally between species and even populations, and is therefore less suitable as a diagnostic criterion for generic delimitation. Two Emmonsia species are reclassified in Emergomyces.

A Fasciclin Protein Is Essential for Laccase-Mediated Selective Phenol Coupling in Sporandol Biosynthesis.

The biaryl scaffold, often showing axial chirality, is a common feature of various fungal natural products. Their biosynthesis requires an oxidative phenol-coupling reaction usually catalyzed by laccases, cytochrome P450 enzymes, or peroxidases. The combination of a laccase and a fasciclin domain-containing (fas) protein is encoded in many biosynthetic gene clusters of biaryls from ascomycetes. However, such phenol-coupling systems including their regio- and stereoselectivity have not been characterized so far. Elucidating the biosynthesis of the antiparasitic binaphthalene sporandol from Chrysosporium merdarium, we demonstrate the combination of a laccase and a fas protein to be crucial for the dimerization reaction. Only the heterologous coproduction of the laccase and the fas protein led to a functional phenol-coupling system, whereas the laccase alone showed no coupling activity. Thus, the laccase/fas protein combination forms an independent group of phenol-coupling enzymes that determines the coupling activity and selectivity of the reaction concurrently and applies to the biosynthesis of many fungal natural products with a biaryl scaffold.

Comparative characterization of extracellular enzymes secreted by Phanerochaete chrysosporium during solid-state and submerged fermentation.

Influence of water content on the expression of lignocellulolytic enzymes by Phanerochaete chrysosporium remains unclear. This work compares the enzyme production profiles of P. chrysosporium during solid-state and submerged fermentation. There were 110 and 64 extracellular carbohydrate-active enzymes identified in solid-state and submerged fermentation respectively, among which 57 enzymes were common to both of the secretomes. P. chrysosporium secreted more cellulases (especially lytic polysaccharide monooxygenase) and hemicellulases during solid-state fermentation while the proportion of enzyme containing carbohydrate-binding module was higher for submerged fermentation. Although its activities were weaker, the enzyme cocktail from submerged fermentation was surprisingly more effective in hydrolysis at low substrate loading. This advantage of enzymes from submerged fermentation was mainly attributed to carbohydrate-binding module because more xylanases bound with substrate at the beginning of hydrolysis. These results reveal the influence of fermentation conditions on enzyme produced by P. chrysosporium for the first time and show the importance of carbohydrate-binding module in the hydrolysis process of lignocellulose.

Chrysosporazines F-M: P-Glycoprotein Inhibitory Phenylpropanoid Piperazines from an Australian Marine Fish Derived Fungus, Chrysosporium sp. CMB-F294.

Chemical analysis of the fungus Chrysosporium sp. CMB-F294 isolated from the gastrointestinal tract of a market-purchased specimen of Mugil mullet yielded eight new alkaloids, belonging to a rare class of phenylpropanoid piperazines. Chrysosporazines F-M (1-8) occur as an equilibrium mixture of acetamide rotamers and feature unprecedented carbocyclic and heterocyclic scaffolds. Structures inclusive of absolute configuration were assigned by detailed spectroscopic analysis, supported by biosynthetic considerations. Structure-activity relationship studies determined that selected chrysosporazines were promising noncytotoxic inhibitors of the multidrug resistance efflux pump P-glycoprotein (P-gp), capable of reversing doxorubicin resistance in P-gp-overexpressing human colon carcinoma cells (SW620 Ad300). Chrysosporazine F (1) was particularly noteworthy, with a 2.5 µM cotreatment inducing a doxorubicin gain in sensitivity (GS 14) > 2-fold that of the positive control verapamil (GS 6.1).

Molecular Diagnosis of Emmonsia-Like Fungi Occurring in Wild Animals.

Using specific primers based on the ribosomal operon, positive DNA amplification was obtained from lungs of 11/215 tested small burrowing animals, both terrestrial and aquatic, and including frozen (n = 4) and formalin-fixed paraffin-embedded (n = 7) samples. The main species detected in Europe in mice, otters and river rats was Emmonsia crescens. Two strains from otters and weasels were Blastomyces parvus. Two Australian wombats revealed the presence of a hitherto unknown species of the geophilic genus Emmonsiellopsis.

Identification and characterization of compounds from Chrysosporium multifidum, a fungus with moderate antimicrobial activity isolated from Hermetia illucens gut microbiota.

The gut microbiota of insects is composed of a wide range of microorganisms which produce bioactive compounds that protect their host from pathogenic attack. In the present study, we isolate and identify the fungus Chrysosporium multifidum from the gut of Hermetia illucens larvae. Extract from C. multifidum culture broth supernatant showed moderate activity against a strain of methicillin-resistant Staphylococcus aureus (MRSA). Bioguided isolation of the extract resulted in the characterization of six α-pyrone derivatives (1-6) and one diketopiperazine (7). Of these compounds, 5,6-dihydro-4-methoxy-6-(1-oxopentyl)-2H-pyran-2-one (4) showed the greatest activity (IC50 = 11.4 ± 0.7 µg/mL and MIC = 62.5 µg/mL) against MRSA.

Impact of the Cultivation Technique on the Production of Secondary Metabolites by Chrysosporium lobatum TM-237-S5, Isolated from the Sponge Acanthella cavernosa.

The fungi Chrysosporium lobatum TM-237-S5 was isolated from the sponge Acanthella cavernosa, collected from the mesophotic coral ecosystem of the Red Sea. The strain was cultivated on a potato dextrose agar (PDA) medium, coupling solid-state fermentation and solid-state extraction (SSF/SSE) with a neutral macroreticular polymeric adsorbent XAD Amberlite resin (AMBERLITE XAD1600N). The SSF/SSE lead to high chemodiversity and productivity compared to classical submerged cultivation. Ten phenalenone related compounds were isolated and fully characterized by one-dimensional and two-dimensional NMR and HRMS. Among them, four were found to be new compounds corresponding to isoconiolactone, (-)-peniciphenalenin F, (+)-8-hydroxyscleroderodin, and (+)-8-hydroxysclerodin. It is concluded that SSF/SSE is a powerful strategy, opening a new era for the exploitation of microbial secondary metabolites.