In vitro activity of eight antifungal drugs against Chaetomiaceae.

The incidence of infections caused by uncommon Chaetomiaceae (Chaetomium and related species) in humans has increased in the recent years. The in vitro activity of eight antifungal drugs (amphotericin B, five azoles, two echinocandins) against 42 morphologically identified Chaetomium strains was determined according to the Clinical and Laboratory Standards Institute (CLSI) guideline. The strains were subsequently identified based on sequences of the internal transcribed spacer 1 and 2 including the intervening 5.8S nrDNA region (ITS) and the partial ß tubulin gene (tub2). Chaetomium globosum (n = 24), was the most frequently isolated species, followed by Amesia atrobrunnea (syn. Chaetomium atrobrunnea, n = 6), Dichotomopilus dolichotrichus (syn. Chaetomium dolichotrichum, n = 2) and Acrophialophora jodhpurensis, Chaetomium coarctatum, C. elatum, C. gracile, C. subaffine, C. tarraconense, C. unguicola, Dichotomopilus sp., Dichotomopilus variostiolatus, Ovatospora brasiliensis (all represented by a single strain). The geometric means of the minimum inhibitory concentrations/minimum effective concentrations (MICs/MECs) of the antifungals across all strains were (in increasing order): micafungin 0.12 µg/ml, itraconazole and posaconazole 0.21 µg/ml, amphotericin B 0.25 µg/ml, voriconazole 0.45 µg/ml, isavuconazole 0.54 µg/ml, caspofungin 2.57 µg/ml, and fluconazole 45.25 µg/ml. Micafungin had the lowest geometric mean followed by amphotericin B which had the largest range against tested isolates. All examined C. globosum strains had similar antifungal susceptibility patterns. Fluconazole and caspofungin could not be considered as an option for treatment of infections caused by Chaetomium and chaetomium-like species. LAY SUMMARY: Infections caused by uncommon fungi such as Chaetomium have increased in the recent years. Chaetomium globosum has been reported from onychomycosis and phaeohyphomycosis. This species often induces superficial infections in immunocompetent patients. The taxonomy of Chaetomium spp. has changed dramatically in the last years. Antifungal treatment is a crucial step for managing these kinds of infections. Therefore, the in vitro activity of eight antifungal drugs against Chaetomium strains was determined and ß-tubulin (tub2) sequencing was applied to identify the strains. Chaetomium globosum was the most frequent species in our dataset. Based on the results of susceptibility testing, micafungin had the lowest geometric mean followed by amphotericin B. Fluconazole and caspofungin cannot be considered a proper treatment option for infections caused by Chaetomium and chaetomium-like species.

Efficient biosynthesis of anticancer polysaccharide by a mutant Chaetomium globosum ALE20 via non-sterilized fermentation.

The sterilization process, due to its immense energy consumption, high facilities investment, and loss of raw materials by caramelization, during industrial production has drawn much attention. In this study, a methanol-resistant mutant strain, Chaetomium globosum ALE20, was obtained following 20 cycles of adaptive laboratory evolution process. The titer of anticancer polysaccharide (GCP-M) from C. globosum ALE20 reached 9.2 g/L with glycerol as sole carbon source using non-sterilized and fed-batch fermentation strategy. This titer represents a 200% increase compared with the 3.3 g/L attained with batch fermentation. The GCP-M monosaccharide was comprised of galactose, glucose, mannose and glucuronic acid, in a molar ratio of 3.83:66.37:3.26:1.95, respectively, and its weight-average molecular weight and polydispersity were 3.796 × 104 Da and 1.060, respectively. This work presents an ideal alternative and safer fermentation process without sterilization, and a useful approach for enhancing industrial production.

In vitro activities of six antifungal agents and their combinations against Chaetomium spp.

PURPOSE: To assess in vitro activities of six antifungal agents (amphotericin B, itraconazole, voriconazole, posaconazole, caspofungin and terbinafine) and the combined effects of eight pairs of them (caspofungin or terbinafine with amphotericin B, itraconazole, voriconazole or posaconazole) against 22 isolates of Chaetomium spp. METHODOLOGY: The broth microdilution method drafted by the Clinical and Laboratory Standards Institute and the checkerboard method were used in this study to evaluate in vitro activities of antifungal drugs both alone and in combination against Chaetomium spp. RESULTS: Amphotericin B and triazoles exhibited lower geometric mean, MIC50 and MIC90 than caspofungin and terbinafine. Besides, all the paired drugs displayed varying degrees of synergism, with the interactions between caspofungin and itraconazole ranking first (86.36 %). CONCLUSION: Our study illustrated varying degrees of synergism between caspofungin or terbinafine and itraconazole, voriconazole, posaconazole or amphotericin B towards Chaetomium spp., which could be a reference for the clinical treatment of Chaetomium spp. infections.

Morphological evolution of various fungal species in the presence and absence of aluminum oxide microparticles: Comparative and quantitative insights into microparticle-enhanced cultivation (MPEC).

The application of microparticle-enhanced cultivation (MPEC) is an attractive method to control mycelial morphology, and thus enhance the production of metabolites and enzymes in the submerged cultivations of filamentous fungi. Unfortunately, most literature data deals with the spore-agglomerating species like aspergilli. Therefore, the detailed quantitative study of the morphological evolution of four different fungal species (Aspergillus terreus, Penicillium rubens, Chaetomium globosum, and Mucor racemosus) based on the digital analysis of microscopic images was presented in this paper. In accordance with the current knowledge, these species exhibit different mechanisms of agglomerates formation. The standard submerged shake flask cultivations (as a reference) and MPEC involving 10 µm aluminum oxide microparticles (6 g·L-1 ) were performed. The morphological parameters, including mean projected area, elongation, roughness, and morphology number were determined for the mycelial objects within the first 24 hr of growth. It occurred that heretofore observed and widely discussed effect of microparticles on fungi, namely the decrease in pellet size, was not observed for the species whose pellet formation mechanism is different from spore agglomeration. In the MPEC, C. globosum developed core-shell pellets, and M. racemosus, a nonagglomerative species, formed the relatively larger, compared to standard cultures, pellets with distinct cores.

Nonsteroidal Anti-inflammatory Drugs (NSAIDS) Inhibit the Growth and Reproduction of Chaetomium globosum and Other Fungi Associated with Water-Damaged Buildings.

Indoor mold due to water damage causes serious human respiratory disorders, and the remediation to homes, schools, and businesses is a major expense. Prevention of mold infestation of building materials would reduce health problems and building remediation costs. Nonsteroidal anti-inflammatory drugs (NSAIDs) inhibit yeasts and a limited number of filamentous fungi. The purpose of this research was to determine the possible inhibitory activity of nonsteroidal anti-inflammatory drugs (NSAIDs) on germination, fungal growth, and reproduction of Chaetomium globosum and other important filamentous fungi that occur in water-damaged buildings. Several NSAIDs were found to inhibit C. globosum germination, growth, and reproduction. The most effective NSAIDs inhibiting C. globosum were ibuprofen, diflunisal, and diclofenac. Fusarium oxysporum, Fusarium solani, Aspergillus niger, and Stachybotrys atra were also tested on the various media with similar results obtained. However, F. oxysporum and A. niger exhibited a higher level of resistance to aspirin and NaSAL when compared to the C. globosum isolates. The inhibition exhibited by NSAIDs was variable depending on growth media and stage of fungal development. These compounds have a great potential of inhibiting fungal growth on building materials such as gypsum board. Formulations of sprays or building materials with NSAID-like chemical treatments may hold promise in reducing mold in homes and buildings.

Developing genetic tools to exploit Chaetomium thermophilum for biochemical analyses of eukaryotic macromolecular assemblies.

We describe a method to genetically manipulate Chaetomium thermophilum, a eukaryotic thermophile, along with various biochemical applications. The transformation method depends on a thermostable endogenous selection marker operating at high temperatures combined with chromosomal integration of target genes. Our technique allows exploiting eukaryotic thermophiles as source for purifying thermostable native macromolecular complexes with an emphasis on the nuclear pore complex, holding great potential for applications in basic science and biotechnology.

Antifungal activity of nano and micro charcoal particle polymers against Paecilomyces variotii, Trichoderma virens and Chaetomium globosum.

This study investigates the antifungal activity of a polymer integrated with nano-porous charcoal particles against Paecilomyces variotii, Chaetomium globosum, Trichoderma virens, which are all filamentous fungi. The charcoal polymers were prepared by combining charcoal powders with plastic resin under a vacuum to form charcoal particle protrusions on the polymer surface. The mycelial growth of P. variotii and T. virens exhibited a reduction of 10 and 30%, respectively, after the conidia were pre-treated with charcoal polymers, and in particular, no mycelial growth was found in C. globosum during 5 days of culture. The adsorption of Ca(2+) into charcoal was suggested to inhibit growth due to the reduction in the flux of calcium ions (Ca(2+)) into the hyphae. In 5 h, about 15 mM of Ca(2+) were removed from CaCl2 solution with 0.2 g/mL of polymers, and the nano-sized pores of the charcoals on the polymer were responsible for the Ca(2+) adsorption.

Epigenetic stimulation of polyketide production in Chaetomium cancroideum by an NAD(+)-dependent HDAC inhibitor.

Exposure of the fungus Chaetomium cancroideum to an NAD(+)-dependent HDAC inhibitor, nicotinamide, enhanced the production of aromatic and branched aliphatic polyketides, which allowed us to isolate new secondary metabolites, chaetophenol G and cancrolides A and B. Their structures were determined using spectroscopic analyses, and their absolute configuration was elucidated by electronic circular dichroism (ECD), vibrational circular dichroism (VCD), and chemical transformations. Biosynthesis of the branched aliphatic polyketide skeletons in cancrolides A and B was evidenced by conducting a feeding experiment using compounds labeled with a (13)C stable isotope.

Photodynamic activity of nanostructured fabrics grafted with xanthene and thiazine dyes against opportunistic fungi.

Fungi are an important class of human pathogens for which considerable research has gone into defeating them. The photodynamic effects of rose bengal (RB), phloxine B (PB), azure A (AA), and toluidine blue O (TBO) dyes to inhibit Aspergillus fumigatus, Aspergillus niger, Trichoderma viride, Penicillium funiculosum, and Chaetomium globosum were investigated grafted to nano- and micro-structured fabrics. Three antifungal tests conducted: broth microdilution test of free dyes, zone of inhibition and quantitative antifungal assays on fabrics grafted with dyes. In the broth microdilution test, free RB displayed the lowest MIC at 32 µM to inhibit visible hyphal growth and germination but the antifungal ability of MIC for other photosensitizers below 63 µM was insignificant. RB and PB showed lower MIC than AA and TBO. In the inhibition zone tests, nanostructured fabrics grafted with RB and PB did not display fungal growth on the surface. Most microstructured fabrics grafted with AA and TBO showed little inhibition. In quantitative antifungal assay, nanostructured fabrics grafted with RB has the largest inhibition rate on T. viride and the lowest inhibition rate on P. funiculosum and the results showed the increasing inhibition rate in the order of AA < TBO < PB < RB.

Organic neem compounds inhibit soft-rot fungal growth and improve the strength of anthracite bricks bound with collagen and lignin for use in iron foundry cupolas.

AIMS: To examine organic neem compounds for their effective growth inhibition of saprotrophic soft-rot fungi on anthracite bricks bound with collagen and lignin for use in iron foundry cupolas as an alternative fuel source. METHODS AND RESULTS: Azadirachtin, crude neem oil (NO), and clarified neem oil extract (CNO) were combined with copper to inhibit the growth of the soft-rot fungus, Chaetomium globosum. A synergistic interaction was observed between CNO and a low dose of copper on nutrient media (two-factor anova with triplicate replication: P < 0·05). Interaction was confirmed on lab-scale collagen-lignin-anthracite briquettes by measuring their unconfined compressive (UC) strength. The effective collagen strength of the briquettes was enhanced by applying CNO to their surface prior to inoculation: the room temperature UC strength of the briquettes was 28 ± 4·6% greater when CNO (0·4 mg cm(-2) ) was surface-applied, and was 43 ± 3·0% greater when CNO plus copper (0·14 µg cm(-2) ) were surface-applied. CONCLUSION: Surface application of CNO and copper synergistically prevents fungal growth on bindered anthracite briquettes and increases their room temperature strength. SIGNIFICANCE AND IMPACT OF THE STUDY: This novel organic fungicidal treatment may increase the storage and performance of anthracite bricks in iron foundries, thereby saving 15-20% of the energy used in conventional coke production.

The peptide toxin amylosin of Bacillus amyloliquefaciens from moisture-damaged buildings is immunotoxic, induces potassium efflux from mammalian cells, and has antimicrobial activity.

Amylosin, a heat-stable channel-forming non-ribosomally synthesized peptide toxin produced by strains of Bacillus amyloliquefaciens isolated from moisture-damaged buildings, is shown in this paper to have immunotoxic and cytotoxic effects on human cells as well as antagonistic effects on microbes. Human macrophages exposed to 50 ng of amylosin ml(-1) secreted high levels of cytokines interleukin-1ß (IL-1ß) and IL-18 within 2 h, indicating activation of the NLRP3 inflammasome, an integral part of the innate immune system. At the same exposure level, expression of IL-1ß and IL-18 mRNA increased. Amylosin caused dose-dependent potassium ion efflux from all tested mammalian cells (human monocytes and keratinocytes and porcine sperm cells) at 1 to 2 µM exposure. Amylosin also inhibited the motility of porcine sperm cells and depolarized the mitochondria of human keratinocytes. Amylosin may thus trigger the activation of the NLRP3 inflammasome and subsequently cytokine release by causing potassium efflux from exposed cells. The results of this study indicate that exposure to amylosin activates the innate immune system, which could offer an explanation for the inflammatory symptoms experienced by occupants of moisture-damaged buildings. In addition, the amylosin-producing B. amyloliquefaciens inhibited the growth of both prokaryotic and eukaryotic indoor microbes, and purified amylosin also had an antimicrobial effect. These antimicrobial effects could make amylosin producers dominant and therefore significant causal agents of health problems in some moisture-damaged sites.

Biological evaluation of endophytic fungus, Chaetomium globosum JN711454, as potential candidate for improving drug discovery.

The main objective of this research work focused on investigating the biological and chemical aspects of endophytic fungus Chaetomium globosum, for pharmaceutical purposes to improve the drug discovery process. The endophytic C. globosum was isolated from healthy leaves of Egyptian medicinal plant Adiantum capillus-veneris collected from Saint Katherine Protectorate, Sinai, Egypt. The identification of C. globosum was on the basis of classical and molecular taxonomy. Gene encoding for 18S rRNA was partially sequenced, submitted to the GenBank and got the accession number JN711454, to resolve the phylogenetic relations with fungal ancestor using phylogenetic tree. To explore the biosynthetic power of endophytic C. globosum JN711454, the fungus was cultivated over five different media, oatmeal, rice, yeast malt glucose, potato dextrose agar (PDA) and Czapek's dox media, for 3 weeks at 30 °C, followed by extraction with different solvents, ethyl acetate (EA), and methanol. The ethyl acetate extract of C. globosum cultivated on PDA medium was the most potent extract. It showed strong antioxidant activity with EC50 11.5 µg/ml, potent anticancer activity with 55 % toxicity toward HepG-2 cells at 100 µg/ml and 66 % cytotoxicity to FGC4 cells at 250 µg/ml, promising butyrylcholinesterase inhibitory activities (>85 %), and moderate antimicrobial and stopped the attachment of HSV-2 virus to VERO cells. The metabolomic profiling of PDA-EA extract using LC-MS revealed the presence of several metabolites to which the observed bioactivities could be attributed. Here we report for the first time inhibitory activity of endophytic C. globosum JN711454 secondary metabolites to butyrylcholinesterase, one of neuro hydrolase enzymes that play a major role in development of Alzheimer's disease.

Method for rapid detection and identification of chaetomium and evaluation of resistance to peracetic acid.

In the beverage industry, peracetic acid has been increasingly used as a disinfectant for the filling machinery and environment due to merits of leaving no residue, it is safe for humans, and its antiseptic effect against fungi and endospores of bacteria. Recently, Chaetomium globosum and Chaetomium funicola were reported resistant to peracetic acid; however, little is known concerning the detail of peracetic acid resistance. Therefore, we assessed the peracetic acid resistance of the species of Chaetomium and related genera under identical conditions and made a thorough observation of the microstructure of their ascospores by transmission electron microscopy. The results of analyses revealed that C. globosum and C. funicola showed the high resistance to peracetic acid (a 1-D antiseptic effect after 900 s and 3-D antiseptic effect after 900 s) and had thick cell walls of ascospores that can impede the action mechanism of peracetic acid. We also developed specific primers to detect the C. globosum clade and identify C. funicola by using PCR to amplify the ß-tubulin gene. PCR with the primer sets designed for C. globosum (Chae 4F/4R) and C. funicola (Cfu 2F/2R) amplified PCR products specific for the C. globosum clade and C. funicola, respectively. PCR with these two primer sets did not detect other fungi involved in food spoilage and environmental contamination. This detection and identification method is rapid and simple, with extremely high specificity.

The application of bioactive compounds from the food industry to control mold growth in indoor waterborne coatings.

Microbial growth in indoor environments creates health problems, especially in people with asthma; approximately 80% of these patients are allergic to mold. Antimicrobial coatings are formulated to generate surfaces that are easy to clean and may also incorporate active agents, commonly called biocides, which inhibit microbial colonization, subsequent growth and bio-deterioration of the substrates. Some research lines seek to replace traditional organometallic and organochlorines biocides with environmentally acceptable ones. The aim of this research was, primarily, to explore the possible application of different compounds used in food industry like preservatives to be used as antimicrobial additives for antimicrobial coatings. Four biocides were tested against two different ambient molds isolated from an interior painted wall (Chaetomium globosum and Alternaria alternate). The selected biocides were zinc salicylate, zinc benzoate, calcium benzoate and potassium sorbate. The resulting paints were subjected to biological and physical tests (viscosity, hiding power, humidity absorption and biocides leaching rate). Bioassays revealed that zinc benzoate and zinc salicylate resulted active against both fungi.

Differential chlorate inhibition of Chaetomium globosum germination, hyphal growth, and perithecia synthesis.

Chaetomium globosum Kunze:Fr is a dermatophytic, dematiaceous fungus that is ubiquitous in soils, grows readily on cellulolytic materials, and is commonly found on water-damaged building materials. Chlorate affects nitrogen metabolism in fungi and is used to study compatibility among anamorphic fungi by inducing nit mutants. The effect of chlorate toxicity on C. globosum was investigated by amending a modified malt extract agar (MEA), oat agar, and carboxymethyl cellulose agar (CMC) with various levels of potassium chlorate (KClO(3)). C. globosum perithecia production was almost completely inhibited (90-100 %) at low levels of KClO(3) (0.1 mM) in amended MEA. Inhibition of perithecia production was also observed on oat agar and CMC at 1 and 10 mM, respectively. However, hyphal growth in MEA was only inhibited 20 % by 0.1-100 mM KClO(3) concentrations. Hyphal growth was never completely inhibited at the highest levels tested (200 mM). Higher levels of KClO(3) were needed on gypsum board to inhibit perithecia synthesis. In additional experiments, KClO(3) did not inhibit C. globosum, Fusarium oxysporum, Aspergillus niger, Penicillum expansum, and airborne fungal spore germination. The various fungal spores were not inhibited by KClO(3) at 1-100 mM levels. These results suggest that C. globosum perithecia synthesis is more sensitive to chlorate toxicity than are hyphal growth and spore germination. This research provides basic information that furthers our understanding about perithecia formation and may help in developing control methods for fungal growth on building materials.

Endophytic Chaetomium globosum enhances maize seedling copper stress tolerance.

This study aims at characterisation of the impact of Chaetomium globosum on copper stress resistance of maize seedlings. Higher levels of copper treatment decreased maize dry weight and induced a marked increase in osmotic solutes, antioxidant enzyme activity and the level of lipid peroxidation. On the other hand, addition of the endophytic C. globosum alleviated the toxic effect of copper on maize growth. The combination of copper sulphate and Chaetomium increased seedling dry weight, osmotic solute content and antioxidant enzyme activity compared to copper sulphate alone, while lipid peroxidation levels were also decreased. The fungal scavenger system might be important for supporting the ability of maize seedlings to resist copper toxicity.

[Effect of PEG stress on plantlets of Chrysanthemum morifolium induced by endophytic botrytis sp. (C1) and Chaetomium globosum (C4)].

The effect of the endophytic fungi Botrytis sp. (C1) or Chaetomium globosum (C4) on the drought resistance of Chrysanthemum morifolium was studied. Ch. morifolium plantlets were inoculated with C1, C4 and cultured in the pots for 60 days, then the plantlets were stressed by 0%, 10%, 20%, 30%, 40% PEG6000 respectively in order to simulate different drought conditions. Biomass, the activities of SOD, POD, PAL, the contents of MDA and soluble protein of each group were determined. The results showed that endophytic fungi groups grew better than the control (without inoculation endophytic fungi). With the increasing of the concentration of PEG6000, the biomass of Ch. morifolium of each groups decreased, while the biomass of fungi groups was significantly higher than that of control, moreover C4 group higher than C1 group. With the concentration of PEG increasing, the content of MDA of each group increased too, while POD activity and soluble protein content of all treatments increased at first and then decreased. SOD activity and PAL activity of the control were increased with the increase of PEG concentration, but SOD activity of the two fungi groups were stable. After been stressed by different concentrations of PEG, MDA content of two fungi groups were always lower than the control, while SOD activity, POD activity, PAL activity and soluble protein content were higher. In conclusion, endophytic fungi can increase the drought resistance of Ch. morifolium.

Antimicrobial/Antimold polymer-grafted starches for recycled cellulose fibers.

In this work, an antimicrobial guanidine polymer (PHGH) was grafted onto starch as a carrier to form branched or grafted chains along the starch backbone. This grafting improved the antimicrobial properties and the adsorption of the starch on recycled cellulose fibers. Similar work was also conducted on bleached sulfite fibers for comparison. The results showed that the starch, grafted with 12 wt% PHGH, adsorbed more on recycled fibers than on sulfite fibers. By applying the antimicrobial-modified starch to recycled or sulfite pulps up to 20 mg/g, both antimicrobial and antimold performances of the papers were improved substantially. Additionally, the PHGH-modified starch increased the tensile index of papers, but decreased the tear index slightly. Scanning electron microscopy (SEM) and atomic force microscopy (AFM) were employed to investigate the morphologic changes of Escherichia coli bacteria and Chaetomium globosum fungi upon exposure to the PHGH-modified starch, thus demonstrating that the antimicrobial mechanism is based on the damage of bacterial membrane.

Evaluating the combined efficacy of polymers with fungicides for protection of museum textiles against fungal deterioration in Egypt.

Fungal deterioration is one of the highest risk factors for damage of historical textile objects in Egypt. This paper represents both a study case about the fungal microflora deteriorating historical textiles in the Egyptian Museum and the Coptic museum in Cairo, and evaluation of the efficacy of several combinations of polymers with fungicides for the reinforcement of textiles and their prevention against fungal deterioration. Both cotton swab technique and biodeteriorated textile part technique were used for isolation of fungi from historical textile objects. The plate method with the manual key was used for identification of fungi. The results show that the most dominant fungi isolated from the tested textile samples belong to Alternaria, Aspergillus, Chaetomium, Penicillium and Trichoderma species. Microbiological testing was used for evaluating the usefulness of the suggested conservation materials (polymers combined with fungicides) in prevention of the fungal deterioration of ancient Egyptian textiles. Textile samples were treated with 4 selected polymers combined with two selected fungicides. Untreated and treated textile samples were deteriorated by 3 selected active fungal strains isolated from ancient Egyptian textiles. This study reports that most of the tested polymers combined with the tested fungicides prevented the fungal deterioration of textiles. Treatment of ancient textiles by suggested polymers combined with the suggested fungicides not only reinforces these textiles, but also prevents fungal deterioration and increases the durability of these textiles. The tested polymers without fungicides reduce the fungal deterioration of textiles but do not prevent it completely.

Onychomycosis by Chaetomium spp.

Onychomycosis is one of the most common infections of the nail. It can be caused by dermatophytes, Candida species and other fungi. Although moulds can cause onychomycosis, they account for a minority of cases, mainly great toenail onychomycosis. We describe a case of an 11-year-old girl who developed an onychomycosis by Chaetomium species in second and third toenails.