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A new, nonglycosylated, serine proteinase capable of hydrolyzing collagen, elastin, and fibrin was isolated from a complex preparation of proteins formed by the micromycete A. ustus 1 under solid-state fermentation. The enzyme had a molecular weight of 33 kDa and a pI of 4.6. The pH-optimum (6.0) and temperature optimum (41°C) of the work of this proteinase were also determined.

Bioactivity-guided isolation of Aspergillus ustus led to the discovery of five new drimane sesquiterpenes, named ustusal A, ustusolate F and G, and ustusoic acid A and B, 1-5 respectively. Structural elucidation of these fungal terpenes relied on 1D and 2D NMR techniques, high-resolution mass spectrometry, and chiroptical properties. Their relative configurations were determined by NMR methods, while the absolute configurations were established using comparative analyses of computed and experimental NMR chemical shifts and ECD spectra. The sesquiterpenes exhibited weak activity against the clinically relevant pathogens vancomycin-resistant Enterococcus faecium and multidrug-resistant Staphylococcus aureus; however, the activity of 5 was drastically enhanced when equal amounts of stromemycin (6), a known metabolite coisolated from the same fraction from A. ustus, was added.

A highly oxygenated phenethyl derivative ustethylin A was isolated from Aspergillus ustus. Gene deletion, isotope labeling, and heterologous expression proved that the phenethyl core structure is assembled from malonyl-CoA by a polyketide synthase harboring a methyltransferase domain. Propionate was converted via acetyl-CoA to malonyl-CoA and incorporated into the molecule. Modifications on the core structure by three different oxidoreductases and one O-methyltransferase lead to the final product, ustethylin A.

One new dihydrobenzofuran derivative (1), known depsipeptide emericellamide A (2), three known drimanes (3–5) and two artifact drimane derivatives (6, 7) were isolated from the marine-derived fungu...

In the current study, new biomasses were prepared from fungi isolates including, Penicillium chrysogenum and Aspergillus ustus and utilized for the removal of heavy metals. During the study, the in...

New antifungal agrochemicals, derived from 2-(benzylsulfonyl)benzothiazole were synthesized by an environmentally friendly method, using water as reaction medium. These compounds were prepared by a one-pot, two-step synthesis, starting from 2-mercaptobenzothiazole and benzyl halides. The potential fungicides were tested against a panel of phytopathogenic fungi, with many of them showing a significant improvement compared to the non-oxidized analogues and the commercial antifungal Captan. The new derivatives 2-((2-chlorobenzyl)sulfonyl)benzo[d]thiazole (4f) and 2-((4-methylbenzyl)sulfonyl)benzo[d]thiazole (4k) presented remarkable properties, being able to inhibit the growth of two resistant moulds (Aspergillus fumigatus and Aspergillus ustus). Both 4f and 4k could be classified as broad-spectrum fungicides, emerging as possible candidates for the control of these moulds, which have negative impact in food production. Fil: Ballari, María Sol. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigaciones en Físico-química de Córdoba. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas. Instituto de Investigaciones en Físico-química de Córdoba; Argentina Fil: Herrera Cano, Natividad Carolina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Ciencia y Tecnología de Alimentos Córdoba. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas. Instituto de Ciencia y Tecnología de Alimentos Córdoba; Argentina. Universidad Nacional de San Juan. Facultad de Ingeniería. Instituto de Biotecnología; Argentina Fil: Wunderlin, Daniel Alberto. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Ciencia y Tecnología de Alimentos Córdoba. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas. Instituto de Ciencia y Tecnología de Alimentos Córdoba; Argentina Fil: Feresin, Gabriela Egly. Universidad Nacional de San Juan. Facultad de Ingeniería. Instituto de Biotecnología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Santiago, Ana Noemi. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigaciones en Físico-química de Córdoba. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas. Instituto de Investigaciones en Físico-química de Córdoba; Argentina

Oxepinamides are fungal oxepine-pyrimidinone-ketopiperazine derivatives. In this study, we elucidated the biosynthetic pathway of oxepinamide D in Aspergillus ustus by gene deletion, heterologous expression, feeding experiments, and enzyme assays. We demonstrated that the cytochrome P450 enzymes catalyzed highly specific and stereoselective oxepin ring formation.

Aspergillus sections Usti and Cavernicolarum are accommodated in the subgenus Nidulantes. In the present study, a polyphasic approach using morphology and multi-gene phylogeny was applied to investigate the taxonomy of these two sections. Based on the phylogenetic analysis, Aspergillus section Usti includes 25 species, which can be assigned to four series: Calidousti, Deflecti, Monodiorum and Usti. Aspergillus sigarelli is newly described in this section and this species was isolated from a cigarette from PR China and belongs to series Calidousti. It is clearly distinct from other members in this series based on ITS, BenA, CaM and RPB2 sequences. Aspergillus section Usti members like A. calidoustus and A. granulosus are important opportunistic pathogens, it is speculative that more pathogenetic species will be found by using polyphasic taxonomy approaches. Aspergillus section Cavernicolarum includes five species, the growth rates on agar media and size and ornamentation of conidia are important characters for differentiating species in section Cavernicolarum.

Highlights • Vermiculite is a new promising carrier for solid-state fermentation of micromycetes. • Aspergillus have higher protease production on vermiculite in comparison with on other carriers. • The mycelium productivity of Aspergillus was higher not only in comparison with SmF, but also in comparison with other carriers under SSF conditions (from 3 to 18 times)., A new method has been developed to increase the productivity of aspergilli - producers of extracellular proteinases based on their cultivation on vermiculite under solid-state fermentation conditions. The productivity of the mycelium Aspergillus ochraceus L-1 and Aspergillus ustus 1 was 3-18 times higher not only in comparison with submerged cultivation, but also in comparison with growth on other carriers studied under solid-state fermentation conditions. Vermiculite can be considered as a new promising carrier for solid-state fermentation of micromycetes.

The Aspergilli of section Usti (group ustus) are represented by over 20 species, of which Aspergillus calidoustus is the most relevant human pathogen. Invasive aspergillosis (IA) caused by these fungi is rare but could represent an emerging issue among the expanding population of patients with long-term immunosuppression receiving antifungal prophylaxis. Clinicians should be aware of this unusual type of IA, which often exhibits distinct clinical features, such as an insidious and prolonged course and a high occurrence of extra-pulmonary manifestations, such as skin/soft tissue or brain lesions. Moreover, these Aspergillus spp. pose a therapeutic challenge because of their decreased susceptibility to azole drugs. In this review, we outline the microbiological and clinical characteristics of IA due to Aspergillus spp. of section Usti and discuss the therapeutic options.

Background Caffeine, theobromine and theophylline are main purine alkaloid in tea. Theophylline is the downstream metabolite and it remains at a very low level in Camellia sinensis. In our previous study, Aspergillus sydowii could convert caffeine into theophylline in solid-state fermentation of pu-erh tea through N-demethylation. In this study, tea-derived fungi caused theophylline degradation in the solid-state fermentation. The purpose of this study is identify and isolate theophylline-degrading fungi and investigate their application in production of methylxanthines with theophylline as feedstock through microbial conversion. Results Seven tea-derived fungi were collected and identified by ITS, β-tubulin and calmodulin gene sequences, Aspergillus ustus, Aspergillus tamarii, Aspergillus niger and A. sydowii associated with solid-state fermentation of pu-erh tea have shown ability to degrade theophylline in liquid culture. Particularly, A. ustus and A. tamarii could degrade theophylline highly significantly (p A. ustus and A. tamarii, respectively. The data from absolute quantification analysis suggested that 3-methylxanthine and xanthine were the main degraded metabolites in A. ustus and A. tamarii, respectively. 129.48 ± 5.81 mg/L of 3-methylxanthine and 159.11 ± 10.8 mg/L of xanthine were produced by A. ustus and A. tamarii in 300 mg/L of theophylline liquid medium, respectively. Conclusions For the first time, we confirmed that isolated A. ustus, A. tamarii degrade theophylline through N-demethylation and oxidation. We were able to biologically produce 3-methylxanthine and xanthine efficiently from theophylline through a new microbial synthesis platform with A. ustus and A. tamarii as appropriate starter strains.

The aim of research was to characterize epiphyte micromycetes observed on variable cotton fibers accessions, to estimate the range of fiber destruction and select cotton, which were the most resistant to fungus damage. The accessions of differently colored Upland Cotton varieties (Gossypium hirsutum L.) evaluated: eleven cotton of natural green, twelve – of brown and eleven of conventional white color. Cotton plants have been grown in Sothern Federal District, RF. The fiber samples for the study were placed into a thermostat in sterile Petri dishes on moistened filter paper in order to stimulate the development of mycelium or sporulation of fungi naturally occurred on fibers. Incubation carried out in a thermostat at a +24-28 °C, humidity of 90-100% and exposed for 28 days. The samples examined with a microscope or binocular magnifier. Aspergillus ustus (Bainier), A. fumigatus Fresen., A. niger v. Tiegh., A. flavus Link, Penicillium aurantiogriseum Dierckx, P. notatum Westling, Rhizopus nigricans Ehrenb. and Alternaria alternata (Fuier) Keissler were detected. Compared with exposed white, accession of green and brown colors were significantly resistant to fungus. The mean of destruction (K) of white cotton varied up to 0.95, but colored accessions not exceeded 0.3 (initial destruction of the surface, not affecting internal fiber’s structure).

Improper disposal and spills of spent engine oil into the environment can result in contamination, which eventually affects humans through the food chain. Mycoremediation is an effective and inexpensive alternative to clean up spent engine oil contamination. In recent work, the potential effectiveness of fungi for degrading spent engine oil was confirmed, with the species identified through molecular identification. Fungi that were able to grow in Bushnell Haas Broth supplied with spent engine oil were identified with the potential to utilize spent engine oil as a carbon source. Six species of fungi namely Penicillium simplicissimum, Aspergillus nidulans, Aspergillus niger, Trichoderma longibrachiatum, Aspergillus ustus, and Aspergillus flavus were successfully identified in this study. Over a course of seven days, P. simplicissimum (21.11 percent) was identified as the most effective fungi in degrading spent engine oil, followed by A. nidulans (17.75 percent), A. niger (15.85 percent), T. longibrachiatum (15.12 percent), A. ustus (15.02 percent), and A. flavus (11.80 percent). As these species of fungi were isolated from the natural environment in Peninsular Malaysia, the potential of using these fungi as mycoremediation of spent engine oil was therefore confirmed.

Seven new ophiobolins (1–5, 12, and 14) along with the 11 known analogues (6–11, 13, 15–18) were isolated from the ethyl acetate extracts of the liquid and solid cultures of the mangrove fungus Aspergillus ustus 094102. The structures including the absolute configurations of the seven new compounds were elucidated by spectroscopic analysis, chemical methods, and quantum ECD calculations. Compounds 4–8 and 11–15 showed cytotoxicities against the G3K, MCF-7, MD-MBA-231, MCF/Adr, A549, and HL-60 human cancer cell lines with the IC50 values ranging from 0.6 to 9.5 μM.

It has been shown that micromycetes Aspergillus ustus 1 and Tolypocladium inflatum k1 secrete proteolytic enzymes that possess high collagenolytic, fibrinolytic, and elastolytic activity. The activity of proteinases hydrolyzing fibrillar proteins, which was determined by the cleavage of azo-collagen, was 122.6 × 10–3EAzc/mL in A. ustus 1 and 69.7 × 10–3EAzc/mL in T. inflatum k1 (EAzc is the amount of azocollagen cleaved in 1 min (μg). The maximum values of activity were observed during submerged cultivation of A. ustus 1 for 4 days and of T. inflatum k1 for 5 days. It has been shown that the maximum of collagenolytic and general proteolytic activity during the cultivation of A. ustus 1 are time-separated, unlike T. inflatum k1, which, presumably, can simplify the procedure for obtaining proteinases active against fibrillar proteins.

Aspergillus spp. cause serious invasive lung infections, and Aspergillus fumigatus is the most commonly encountered clinically significant species. Voriconazole is considered to be the drug of choice for treating A. fumigatus infections; however, rising resistance rates have been reported. We evaluated a matrix-assisted laser desorption ionization–time of flight mass spectrometry (MALDI-TOF MS)-based method for the differentiation between wild-type and non-wild-type isolates of 20 Aspergillus spp. (including 2 isolates of Aspergillus ustus and 1 of Aspergillus calidoustus that were used as controls due their intrinsic low azole susceptibility with respect to the in vitro response to voriconazole). At 30 and 48 h of incubation, there was complete agreement between Cyp51A sequence analysis, broth microdilution, and MALDI-TOF MS classification of isolates as wild type or non-wild type. In this proof-of-concept study, we demonstrated that MALDI-TOF MS can be used to accurately detect A. fumigatus strains with reduced voriconazole susceptibility. However, rather than proving to be a rapid and simple method for antifungal susceptibility testing, this particular MS-based method showed no benefit over conventional testing methods.

Made available in DSpace on 2020-12-10T19:39:46Z (GMT). No. of bitstreams: 0 Previous issue date: 2019-01-01. Added 1 bitstream(s) on 2021-07-15T14:37:10Z : No. of bitstreams: 1 S0001-37652019000500601.pdf: 975738 bytes, checksum: 4ec2cf4858b8200fba6761772dcd422f (MD5) Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) Isolation and screening of new fungal strains from extreme and understudied environments. such as caves, is a promising approach to find higher yields enzyme producers. Cellulolytic fungal strains isolated from a Brazilian cave were evaluated for their enzymatic production after submerged (SmF) and solid-state fermentation (SSF). After SmF, three strains were selected for their high enzymatic activities: Aspergillus ustus for endoglucanase (4.76 U/mg), Talaromyces bruneus for beta-glucosidase (11.71 U/mg) and Aspergillus sp. (CBMAI 1926) for total cellulase (1.70 U/mg). After SSF, these strains, showed better yields compared to the reference strain Aspergillus niger 3T5B8. Aspergillus sp. (CBMAI 1926) stood out as a new species that expressed activity of total cellulases (0.10 U/mg) and low protein concentration (0.44 mg/mL). In conclusion, these isolated strains have a more efficient and promising cellulolytic enzyme complex that can be used in fermentation and saccharification processes with a lower protein concentration and a higher enzymatic activity than the reference strain. Therefore, beside the new genetic material characterized, our study highlights the benefits of cave extreme environments exploitation to find new potentially valuable strains. Univ Fed Sao Carlos, Dept Ecol & Biol Evolut, Programa Posgrad Ecol & Recursos Nat, Rodovia Washington Luiz,Km 235, BR-13565905 Sao Carlos, SP, Brazil Univ Estado Sao Paulo Julio de Mesquita Filho, Inst Biociencias Rio Clara, Dept Bioquim & Microbiol, Av 24A,1515, BR-13506900 Rio Claro, SP, Brazil Empresa Brasileira Pesquisa Agr, Embrapa Instrumentacao, Rua 15 Novetnbro 1452, BR-13560970 Sao Carlos, SP, Brazil Univ Estado Sao Paulo Julio de Mesquita Filho, Inst Biociencias Rio Clara, Dept Bioquim & Microbiol, Av 24A,1515, BR-13506900 Rio Claro, SP, Brazil FAPESP: FAPESP 2015/24763-9

Two new austalide meroterpenoids, named austalides V and W (1 and 2), were isolated from the fungus Aspergillus ustus VKM F-4692. Their structures were elucidated by extensive spectroscopic analysis and by comparison with related known compounds. The main structural feature of both compounds is a tetrahydrofuranyl ring (G), a structural fragment, first found in austalides. Austalides V (1) and W (2) were able to inhibit the propagation of prostate and bladder cancer cells; this biologic activity is possibly related to the inhibition of a number of key pathways regulating cell growth and migration.

Interaction between indigenous fungal strains in preventing ochratoxin A (OTA) production by Penicillium verrucosum and Penicillium nordicum was studied in 100 mL of Czapek yeast autolysate (CYA) medium in a 250-mL “U” shaped culture vessel in one end for 3 days. At the end of incubation period, test fungi inoculated and incubated at 27 ± 2 °C for another 14 days to study the inhibition of OTA production was estimated by high performance liquid chromatography (HPLC). Total inhibition of OTA production was recorded with Aspergillus fumigatus , Aspergillus flavus , Aspergillus niger , Aspergillus ustus , Fusarium culmorum , Fusarium graminarium , Fusarium proliferatum , Penicillium chrysogenum , Penicillium expansum and Trichoderma viridae . A significant correlation coefficient ( r ) on growth (0.493, P ≤ 0.0003) and OTA production (0.785, P ≤ 0.0001) was observed between the tested Penicillium species and other co-existing fungi. In conclusion, the present investigation revealed that those indigenous fungi are necessary to minimize potential losses to the poultry farmer and toxicological hazards to the consumer as biological control agent in different foods and feeds.

Two new sesterterpenoids, terretonins H ( 1 ) and I ( 2 ), together with two known compounds, strobilactone A ( 3 ) and cerebroside D ( 4 ), were isolated from the lipophilic extract of the marine-derived fungus Aspergillus ustus KMM 4664. The structures of compounds 1 and 2 were determined based on spectroscopic methods and confirmed by X-ray crystallographic analysis of terretonin H. All compounds were tested for cytotoxic and embryotoxic effects using sperm and developing embryos of the sea urchin Strongylocentrotus intermedius and human cancer cells.

In the indoor environment, people are exposed to several fungal species. Evident dampness is associated with increased respiratory symptoms. To examine the immune responses associated with fungal exposure, mice are often exposed to a single species grown on an agar medium. The aim of this study was to develop an inhalation exposure system to be able to examine responses in mice exposed to mixed fungal species aerosolized from fungus-infested building materials. Indoor airborne fungi were sampled and cultivated on gypsum boards. Aerosols were characterized and compared with aerosols in homes. Aerosols containing 10 7 CFU of fungi/m 3 air were generated repeatedly from fungus-infested gypsum boards in a mouse exposure chamber. Aerosols contained Aspergillus nidulans , Aspergillus niger , Aspergillus ustus , Aspergillus versicolor , Chaetomium globosum , Cladosporium herbarum , Penicillium brevicompactum , Penicillium camemberti , Penicillium chrysogenum , Penicillium commune , Penicillium glabrum , Penicillium olsonii , Penicillium rugulosum , Stachybotrys chartarum , and Wallemia sebi . They were all among the most abundant airborne species identified in 28 homes. Nine species from gypsum boards and 11 species in the homes are associated with water damage. Most fungi were present as single spores, but chains and clusters of different species and fragments were also present. The variation in exposure level during the 60 min of aerosol generation was similar to the variation measured in homes. Through aerosolization of fungi from the indoor environment, cultured on gypsum boards, it was possible to generate realistic aerosols in terms of species composition, concentration, and particle sizes. The inhalation-exposure system can be used to study responses to indoor fungi associated with water damage and the importance of fungal species composition.

It was shown that extracellular proteinases produced by the strains of micromycetes A. ochraceus L-1 and A. ustus 1 differ by the activity at various pH as well as by the intensity of the effect on fibrillar proteins. It was revealed that the proteinases of A. ochraceus L-1 demonstrated maximum activity during the growth of the producer in the nitrate-free growth medium (the pH of enzyme reaction was 8.0), whereas those of A. ustus 1 showed maximal activity during the growth of the micromycete in the medium containing sodium nitrate (the pH of enzyme reaction was 6.0). Values of specific fibrinolytic and collagenolytic activities of A. ochraceus L-1 were 2.2 and 1.6 times higher than those of A. ustus 1. At the same time, A. ustus 1 showed very low values of total proteolytic (caseinolytic) activity and had a high ratio of fibrinolytic activity to total proteolytic (caseinolytic) activity (6.92). It makes the strain a promising producer of proteinases, which hydrolyze fibrin and collagen.

The purpose of research is increase of honey productivity bee colonies. The studies were conducted in the early spring in March, before the departure of the insect from the hive after hibernation. Bee colonies of the experimental group treated with ozone for families for 7 consecutive days at 2 hours per day when the ozone concentration of 6 mg . Bee colonies of the control group not treated with ozone. The results showed the presence of different species of microorganisms in the intestinal tract of bees in the control group, the quantitative indicators of the content of the polymorphic bacteria, staphylococci, streptococci, pseudomonas, fungi is quite large. Also, the experiments proved devastating effect of ozonation for bacteria groups, coliforms, pseudomonas, fungi of the genus aspergillus niger in the experimental group. With regard to the fungi penicillium glaucum and aspergillus ustus, ozone caused a slight decrease in their concentration in the intestinal tract of bees. Therefore, the use of ozone as a means of prevention and treatment of infectious diseases of bees can be justified as it creates the most favorable conditions for further development of bee colonies. However, the fact that the number of fungi after ozonation in the intestinal tract of bees increased, speaks about the resistance of these microorganisms to ozone, and therefore, for suppression of fungal flora, ozonation needs to be done either longer or use a higher concentration of ozone.

Breakthrough invasive mold infections (IMIs) that occur during posaconazole or voriconazole prophylaxis are rare complications for which epidemiological data are lacking. This retrospective analysis comparing 24 microbiologically documented breakthrough with 66 nonbreakthrough IMIs shows a shift towards non-Aspergillus molds with a significantly increased proportion of rare multidrug-resistant molds.

Invasive fungal infections due to Aspergillus calidoustus with decreased azole susceptibility are emerging in the setting of azole prophylaxis and are associated with poor outcomes. We assessed the in vitro activity of antifungal drugs used alone or in combinations against A. calidoustus and found a synergistic effect between voriconazole and terbinafine at concentrations within the therapeutic range.

Objectives To investigate the in vitro activity of olorofim (F901318), a novel broad-spectrum antifungal agent, against 150 strains belonging to 16 different cryptic species of Aspergillus by EUCAST and CLSI methodologies. Methods Olorofim, amphotericin B, micafungin, posaconazole and voriconazole were tested against cryptic species belonging to Aspergillus fumigatus complex (n = 57), Aspergillus ustus complex (n = 25), Aspergillus niger complex (n = 20), Aspergillus flavus complex (n = 20), Aspergillus circumdati complex (n = 15) and Aspergillus terreus complex (n = 13) using EUCAST and CLSI methodologies for broth microdilution susceptibility testing of antifungal agents. Results Olorofim was the only drug with activity against all cryptic species of Aspergillus tested, including the multiresistant species Aspergillus lentulus, Aspergillus fumigatiaffinis and Aspergillus calidoustus. Geometric means of MICs for olorofim were lower (0.017, 0.015 and 0.098 mg/L, respectively, for EUCAST; and 0.015, 0.015 and 0.048 mg/L, respectively, for CLSI) than for amphotericin B (4.438, 12.699 and 0.554 mg/L, respectively, for EUCAST; and 0.758, 1.320 and 0.447 mg/L, respectively, for CLSI), voriconazole (2.549, 2.297 and 5.856 mg/L, respectively, for EUCAST; and 2.071, 1.741 and 5.657 mg/L, respectively, for CLSI) and posaconazole (0.307, 0.308 and 12.996 mg/L, respectively, for EUCAST; and 0.391, 0.215 and 9.514 mg/L, respectively, for CLSI). Conclusions Olorofim shows encouraging in vitro activity against cryptic species of Aspergillus that can be hard to treat with current antifungal therapies. Further studies are warranted in order to assess its efficacy.

In the present study, we presented the complete mitochondrial genome of Aspergillus sp. The complete mitochondrial genome of Aspergillus sp. was composed of circular DNA molecules, with a total length of 31,374 bp. The base composition of this mitochondrial genome is as follows: A (36.25%), T (38.09%), G (14.55%), and C (11.10%). The mitochondrial genome contains 18 protein-coding genes, 2 ribosomal RNA genes (rRNA), and 31 transfer RNA genes. The taxonomic status of the Aspergillus sp. mitochondrial genome exhibits a closest relationship with Aspergillus ustus and A. flavus.

Microscopic fungi were isolated from different materials including muscles, bones, skin and funeral clothes from the mummified human remains of three members of the Kuffner's family and from the surrounding air environments. Their hydrolytic abilities such as cellulolytic, lipolytic, and proteolytic-keratinolytic were also assessed. The most isolated fungi, from human remains, belonged mainly to the species of Aspergillus (Aspergillus candidus, Aspergillus calidoustus, Aspergillus fumigatus, Aspergillus niger, Aspergillus sydowii, Aspergillus terreus, Aspergillus ustus, Aspergillus venenatus, Aspergillus versicolor, Aspergillus westerdijkiae) and Penicillium (Penicillium chrysogenum, Penicillium commune, Penicillium crustosum, Penicillium griseofulvum, Penicillium hordei, Penicillium polonicum). Aspergilli and penicillia were the predominant actors also in the air samples, but also many strains belonging to the Rhizopus group were isolated as well. Several fungi exhibited different hydrolytic ability, the most active isolated from human remains belonged to the species A. candidus, A. westerdijkiae, Coprinellus xanthothrix, P. chrysogenum, P. commune, P. griseofulvum and Scopulariopsis brevicaulis. The species recovered from the air displayed stronger deterioration characteristics as compared to human samples. This study can be considered one of few investigations focused on mummified human remains conserved in this kind of the environment.

The growth and survival of some fungi associated with composted and pretreated sawdust particularly for mushroom cultivation were investigated on two growth media; Cooke’s and Oxytetracycline Glucose Yeast Extract (OGYE). Some fungi were isolated during the composting of sawdust over a period of 28 days as well as after pretreatment with gamma irradiation doses of 5, 10, 15, 20, 25 and 32 kGy and moist heat of 100±2oC for 2.5 hours. Fungal counts ranged 4.725.77 log10 CFU/g and 3.44.1 log10 CFU/g respectively for both media. Both pretreatment methods effectively reduced (p

Objectives Invasive mould infections are associated with a high mortality rate and the emergence of MDR moulds is of particular concern. Calcineurin and its chaperone, the heat shock protein 90 (Hsp90), represent an important pathway for fungal virulence that can be targeted at different levels. We investigated the antifungal activity of compounds directly or indirectly targeting the Hsp90-calcineurin axis against different mould species. Methods The in vitro antifungal activity of the anticalcineurin drug FK506 (tacrolimus), the Hsp90 inhibitor geldanamycin, the lysine deacetylase inhibitor trichostatin A and the Hsp70 inhibitor pifithrin-μ was assessed by the standard broth dilution method against 62 clinical isolates of Aspergillus spp. and non-Aspergillus moulds (Mucoromycotina, Fusarium spp., Scedosporium spp., Purpureocillium/Paecilomyces spp. and Scopulariopsis spp.) Results FK506 had variable antifungal activity against different Aspergillus spp. and was particularly active against Mucor spp. Geldanamycin had moderate antifungal activity against Fusarium spp. and Paecilomyces variotii. Importantly, trichostatin A had good activity against the triazole-resistant Aspergillus ustus and the amphotericin B-resistant Aspergillus terreus as well as the MDR Scedosporium prolificans. Moreover, trichostatin A exhibited synergistic interactions with caspofungin against A. ustus and with geldanamycin against Rhizopus spp. for which none of the other agents showed activity. Pifithrin-μ exhibited little antifungal activity. Conclusions Targeting the Hsp90-calcineurin axis at different levels resulted in distinct patterns of susceptibility among different fungal species. Lysine deacetylase inhibition may represent a promising novel antifungal strategy against emerging resistant moulds.

Aeromycoflora in relation to occurrence and prevalence, their seasonal variation and distribution of fungal spores in air was studied from October, 2010 to September, 2011. During the study a total of 31 fungal species belonging to 18 genera of fungi were isolated and identified. Total fungal colony count was 2308. The highest occurrence and prevalence of fungi was contributed by Aspergillus niger 1075 colonies, Penicillium spp. 582 colonies, Aspergillus fumigatus 167, A. flavus 112 colonies, Rhizopus sp. 121 colonies, Candida spp. 39 colonies, Chladosporium spp. 36 colonies, Syncephalestrum sp. 25 colonies, and Fusarium spp. 14 colonies. Lowest colony countwas recorded one, in Aspergillus ustus, Drechslera sp., Nigrospora sp. and Ulocladium sp. Maximum fungal pollutants were found 339 in July, 2010 at temperature 27.12°C and moisture 84.2% during the wet and rainy days and declined in January, 2011 when temperature and moisture label decreased to 20.26°C and 59.5%, respectively. Highest colony density was recorded in July, 2011, it was moderate in February and March, 2011. Lowest colony density was recorded in January and May, 2011 DOI: http://dx.doi.org/10.3329/jbas.v38i1.20211 Journal of Bangladesh Academy of Sciences, Vol. 38, No. 1, 49-59, 2014

One new ergosteroid derivative, isocyathisterol (1), and eight known compounds (2–9) were isolated from the culture of an algicolous strain (cf-42) of Aspergillus ustus obtained from the fresh tissue of marine green alga Codium fragile. The structure and absolute configuration of 1 were unequivocally identified by using NMR and mass spectroscopic methods as well as quantum chemical calculations. Compound 1 exhibited weak antibacterial activity.

The fungus Aspergillus ustus was isolated from external ear inflammation. This fungus rarely infects human because it infects immunocompromised patients causing cutaneous, endocarditis and pneumonia infections. It is also a pathogenic plant and contaminating the crops. The effect of alcoholic extract of leaves of Achillea millefolium (yarrow), Salvia officinalis (sage) and Cassia spp.(seena), was tested by using four concentrations for each extract 5,10,15,20 mg/ml. Results were compared with the antifungal ketoconazole and with the biocontrol agents Trichoderma viride and Trichoderma harzianum. It is noticed that the highest percentage of inhibition was at concentration 20mg/ml for all the plant extracts used for inhibition percentage for A. millefolium, S. officinalis and Cassia spp.were 56.6%, 83.3% and 100% respectively. For ketoconazole the percentage of inhibition was 100% for all concentrations used in this study. The biocontrol agents T. harzianum and T. viride proved to have a high antagonistic ability against the pathogen. This proves that the alcoholic leaves extracts of these plants were more inhibitory for the fungus in addition to biocontrol agents.

Objectives Invasive fungal diseases (IFDs) are life-threatening events in patients with haematologic disorders, and the spectrum of the aetiological pathogens continues to expand. This study aimed to evaluate the clinical utility of a panfungal polymerase chain reaction (PCR) assay for the management of IFDs in such patients. Methods We prospectively analysed 273 consecutive blood samples from 64 risk episodes in 51 patients with haematologic disorders at high risk for IFD who were treated at our hospital between April 2007 and October 2010. Results PCR-positive results were obtained in 18 of 64 risk episodes (35.3%). IFD was documented in 14 episodes (21.9%, 9 probable IFDs and 5 possible IFDs) according to the revised criteria of the European Organization for Research and Treatment of Cancer/Mycoses Study Group. PCR was positive in all of these 14 episodes, and in 4 of the 50 episodes with no IFD category. Sensitivity, specificity, positive predictive value, and negative predictive value of our assay were 100%, 92%, 78% and 100% respectively. A considerable number of fungi (44.4%) that are less common than Aspergillus and Candida species were positive by PCR. Molecular diagnoses of Cunninghamella species, Aspergillus ustus, Fusarium species, Scedosporium apiospermum, Rhodotorula species and Rhizopus species were beneficial in selecting suitable treatments. Conclusions Our panfungal PCR approach allows for the highly sensitive and specific detection and identification of a wide spectrum of fungal pathogens, which provides indispensable information for managing IFDs, especially refractory or breakthrough IFDs during antifungal therapy in high-risk patients with haematologic disorders.

One new meroterpene, 1,2-dihydroterretonin F (2), five new sesterterpenes, (6α)-21-deoxyophiobolin G (3), (6α)-16,17-dihydro-21-deoxyophiobolin G (4), ophiobolin U (5), ophiobolin V (6), and ophiobolin W (7), two new sesquiterpenes, (6-strobilactone-B) esters of (E,E)-6,7-epoxy-2,4-octadienoic acids (13 and 14), and twelve known terpenes (1, 8–12, and 15–20) were isolated from Aspergillus ustus, a fungus from the fresh tissue of marine green alga Codium fragile. Their structures and absolute configurations were identified by NMR and mass spectroscopic methods as well as quantum chemical calculations. Some of the isolates exhibited antibacterial activity and brine shrimp toxicity.

Aspergillus calidoustus, previously classified as Aspergillus ustus, is an emerging pathogen in immunocompromised persons. We describe four recent cases of A. calidoustus and review 37 additional cases of A. calidoustus (n = 8) or A. ustus (n = 29) published through June 2016. Twenty (49%) cases occurred in patients with hematologic malignancy and/or receipt of hematopoietic cell transplantation, and 13 (32%) occurred in solid organ transplant recipients. Antifungal susceptibility was reported in 49% of cases and in 42% treatment failed. Overall mortality was 66% and, where reported, attributable mortality was 30%. A. calidoustus infection is associated with a high mortality rate and frequently displays in vitro antifungal resistance.

Terpenoids are the most diverse and abundant natural products among which sesterterpenes account for less than 2%, with very few reports on their biosynthesis. Ophiobolins are tricyclic 5–8–5 ring sesterterpenes with potential pharmaceutical application. Aspergillus ustus 094102 from mangrove rizhosphere produces ophiobolin and other terpenes. We obtained five gene cluster knockout mutants, with altered ophiobolin yield using genome sequencing and in silico analysis, combined with in vivo genetic manipulation. Involvement of the five gene clusters in ophiobolin synthesis was confirmed by investigation of the five key terpene synthesis relevant enzymes in each gene cluster, either by gene deletion and complementation or in vitro verification of protein function. The results demonstrate that ophiobolin skeleton biosynthesis involves five gene clusters, which are responsible for C15, C20, C25, and C30 terpenoid biosynthesis.

Summary Mycetoma is a chronic granulomatous, subcutaneous disease endemic in tropical and subtropical countries. It is currently a health problem in rural areas of Africa, Asia and South America. Nine cases of mycetoma were analysed in a retrospective study. All isolates were identified by morphological features. The level of species identification was reached by molecular tools. Definitive identification of fungi was performed using sequence analysis of the ITS of the ribosomal DNA region and the ribosomal large-subunit D1/D2. Identification of actinomycetes was accomplished by the 16S rRNA gene sequence. Six unusual clinical isolates were identified: Aspergillus ustus, Cyphellophora oxyspora, Exophiala oligosperma, Madurella pseudomycetomatis, Nocardia farcinica and Nocardia wallacei. The prevalence of mycetoma in Venezuela remains unknown. This study represents the first report in the literature of mycetoma caused by unusual pathogens identified by molecular techniques.

Two new species of Aspergillus that are clearly distinct from all known species in section Usti were revealed during a study of microfungal communities in Spanish caves. The novel species identified in this study and additional species of Aspergillus section Usti are associated with places and substrates related to human activities in caves. Novel species are described using a polyphasic approach that combines data from four loci (ITS, benA, caM and rpb2), morphology and basic chemical and physiological analyses. Aspergillus thesauricus sp. nov. was isolated from various substrates, including the decaying organic matter, cave air and cave sediment of the Cueva del Tesoro Cave (the Treasure cave). The species is represented by twelve isolates and is most closely related to the recently described A. germanicus. Aspergillus baeticus sp. nov. was isolated from the cave sediment in the Gruta de las Maravillas Cave (the Grotto of the Marvels) and is represented by two isolates. Additional isolate was found in the Cueva del Tesoro Cave and in the Demänovska Peace Cave (Slovakia), suggesting a potentially wide distribution of this microorganism. The species is related to A. ustus and A. pseudoustus. Both species are unable to grow at 37 °C, and a weakly positive, yellow Ehrlich reaction was observed in A. thesauricus. Unique morphological features alone are sufficient to distinguish both species from related taxa., This study was supported by the Research Plan of the ISB BC AS CR, v.v.i., presented results were obtained within bilateral projects between AS CR and CSIC “Fungal diversity in Spanish caves with rock art paintings” and “Early detection of fungal outbreaks in Andalusian caves”. Molecular genetics analyses were supported by projects MSM 6007665801, AV0Z50200510 and GAUK 607812.

Chemical examination of the fungus Aspergillus ustus isolated from the Mediterranean sponge Suberites domuncula yielded the five new ophiobolin-type sesterterpenoids 1–5 and the two new pyrrolidine alkaloids 6 and 7, together with the known compound aurantiamine and cerebroside D. The structures of the new compounds were unambiguously elucidated on the basis of extensive spectroscopic-data analysis (1D- and 2D-NMR, MS, and UV) and comparison with literature data. All compounds were evaluated for their cytotoxicity against murine lymphoma cell line L5178Y.

The majority of mycotoxins produced by Aspergillus fungi are immunosuppressive agents, and their cytotoxicity may impair defense mechanisms in humans. The objective of the study was evaluation of the cytotoxicity of fungi isolated from an environment where inpatients with impaired immunity were present. The materials comprised 57 fungal strains: Aspergillus fumigatus, Aspergillus niger: Aspergillus ochraceus, Aspergillus flavus, Aspergillus versicolor and Aspergillus ustus isolated from hospital rooms in Cracow. The cytotoxicity of all the strains was evaluated using the MTT test (3-(4,5-dimethylthiazol-2-yl) 2,5 diphenyltetrazolium bromide). To emphasize the differences in cytotoxicity among the particular strains, variance analysis (ANOVA) and Tukey's difference test were used. Out of 57 Aspergillus strains tested, 48 (84%) turned out to be cytotoxic. The cytyotoxicity was high (+++) in 21 strains, mainly in A. fumigatus. The least cytotoxic were A. niger fungi, this being statistically significant (p

Five new drimane sesquiterpenes (1-5) together with 14 known analogues (6-19) were isolated from laboratory cultures of a mangrove-derived fungus Aspergillus ustus. Their structures were established by spectroscopic methods and antitumor activities were evaluated by sulforhodamine B (SRB) and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) methods.

This review highlights recent findings of our group on bioactive marine natural products isolated from marine sponges and marine derived fungi. The activated chemical defence of the Mediterranean sponge Aplysina aerophoba is introduced as an example of a dynamic response of marine sponges to wounding. Following tissue disrupture preformed brominated isoxazoline alkaloids are enzymatically cleaved and thereby give rise to aeroplysinin-1 which is believed to protect sponges from invasion of pathogenic bacteria. A preliminary characterization of the membrane bound enzyme(s) involved in the cleavage reaction is presented. Bromotyrosine derived, oxime group bearing peptides, the so called bastadins, obtained from the sponge Ianthella basta and some of their synthetic derivatives were shown to exhibit pronounced antifouling activity against larvae of the barnacle Balanus improvisus. The antifouling activity could be traced to the oxime group as an important pharmacophore that was also found to be present in other sponge derived natural products exhibiting antifouling activity. Marine derived fungi that reside within invertebrates such as sponges or inside Mangrove plants are emerging as a new source of bioactive metabolites as demonstrated for Aspergillus ustus and Alternaria sp. that were isolated from the sponge Suberites domuncula or from the Mangrove plant Sonneratia alba, respectively. The former fungus yielded new moderately cytotoxic sesquiterpenoids of the drimane type whereas the latter was found to produce polyketides such as alternariol that exhibited strong and selective inhibitory activity against several protein kinases, for instance Aurora A and B which are targets for anticancer chemotherapy.

Marine endosymbiotic fungi Aspergillus ustus (MSF3) which produce high yield of biosurfactant was isolated from the marine sponge Fasciospongia cavernosa collected from the peninsular coast of India. Maximum production of biosurfactant was obtained in Sabouraud dextrose broth. The optimized bioprocess conditions for the maximum production was pH 7.0, temperature 20 degrees C, salt concentration 3%, glucose and yeast extract as carbon source and nitrogen sources respectively. The response surface methodology based analysis of carbon and nitrogen ratio revealed that the carbon source can increase the biosurfactant yield. The biosurfactant produced by MSF3 was partially characterized as glycolipoprotein based on the estimation of macromolecules and TLC analysis. The partially purified biosurfactant showed broad spectrum of antimicrobial activity. The strain MSF3 can be used for the microbially enhanced oil recovery process.

Zusammenfassung Da das Absterben kleiner Termiten-Kolonien, besonders nach einem Umsetzen in neue Umgebung, mit der Wirkung von Schimmelpilzen in Zusammenhang gebracht wurde, andererseits gewisse Schimmelpilzarten als zusatzliche Nahrung fur die Termiten von Wert sind, wurde der Einflus von rund 20 Ascomycetes-Arten und rund 10 Fungi imperfecti, zum Teil mit mehreren (bis zu 9) Stammen, so das insgesamt uber 60 Pilze benutzt wurden, auf Kalotermes flavicollis (Fabr.), Heterotermes indicola (Wasmann), Reticulitermes lucifugus (Rossi) var. santonensis Feytaud und Nasutitermes ephratae (Holmgren) gepruft. Dabei wurden Sporen, Mycel oder beides an einheitliche Gruppen der Termiten bei gleichbleibenden Temperatur- und Feuchtigkeits-bedingungen verfuttert. Zum Vergleich dienten entsprechende Gruppen ohne jede Nahrung oder mit Filtrierpapier-Futterung. Die Wirkung der einzelnen Schimmelpilzarten und -stamme auf die Termiten reicht von starker Giftigkeit, die zum Absterben aller Tiere im Laufe weniger Tage fuhren kann, bis zu Nutzlichkeit als Nahrungsstoff, bei dem die Tiere langer am Leben bleiben als bei Filtrierpapier-Futterung. Solche Unterschiede wurden nicht nur bei verschiedenen Pilzarten, sondern auch bei den untersuchten Stammen einer Art gefunden. Die verwendeten Termitenarten reagierten nur bei einem Teil der Pilze gleichsinnig. In vielen Fallen verhielten sie sich verschieden. Trichoderma-Arten wirken meist giftig auf Termiten. Aspergillus flavus enthalt sehr giftige, indifferente und ausgesprochen forderliche Stamme. Andere Aspergillus- und mehrere Penicillium-Arten zeigten sehr verschiedenen Einflus. Ausgesprochen gunstig verhielten sich die gepruften Stamme von Aspergillus ustus, Alternaria humicola, Chaetomium-Arten, Dactylium fusarioides, Memnoniella echinata und Trichurus spec., also besonders Cellulose abbauende sogenannte ”Moderfaule„-Pilze. Die Ursachen der verschiedenen Giftigkeit und des unterschiedlichen Verhaltens der Termitenarten sollten durch weitere Versuche, bei denen Toxin-und Antibiotica-Wirkung und die Symbionten-Zusammensetzung bei den Termitenarten zu beachten waren, erforscht werden. Summary The dying of small termite colonies, especially after transfer into a new environment, has been attributed to the action of mould fungi; but on the other hand certain moulds are useful as additional food for the termites. Therefore the influence of about 30 species of Ascomycetes and some Fungiimperfecti, partly represented by several (up to 9) strains, altogether of more than 60 fungus cultures, on Kalotermes flavicollis (Fabr.), Heterotermes indicola (Wasmann), Reticulitermes lucifugus (Rossi) var. santonensis Feytaud, and Nasutitermes ephratae (Holmgren) was tested. Uniform groups of termites were fed with spores, mycelium, or both combined, under constant conditions of temperature and moisture. Equivalent groups, kept without any food or on filter paper, were used as controls. The effect of the individual species and strains of moulds on the termites ranges from strong toxicity, causing death of all specimens within a few days, to usefulness as a nutriment which effects longer survival of the termites than filter paper. This variation was not only stated among different species of fungi, but also among the tested strains of one species. The termite species used reacted only with part of the fungi equally. In many cases they showed different behaviour. Trichoderma species are mostly toxic towards termites. Aspergillus flavus has highly toxic, indifferent, and markedly favourable strains. Other species of Aspergillus and several species of Penicillium varied considerably in their effect. Markedly favourable influence was stated with the tested strains of Aspergillus ustus, Alternaria humicola, Chaetomium spec., Dactylium fusarioides, Memnoniella echinata, and Trichurus spec.; most are especially cellulose-decomposing species, so-called ”soft-rot„ fungi. The causes of the varying toxicity and of the different behaviour of termite species should be investigated by further tests, taking the effect of toxins and antibiotics and the composition of symbionts with the respective termites into consideration.

Seven new drimane sesquiterpenoids (1-3, 6-9), along with the known compounds deoxyuvidin B (4), strobilactone B (5), and RES-1149-2 (10), were obtained from cultures of the fungus Aspergillus ustus, which was isolated from the marine sponge Suberites domuncula. Their structures were established by means of spectroscopic analyses including one- and two-dimensional NMR spectroscopy and high-resolution MS. Compounds 6, 7, and 10 showed cytotoxic activity against a panel of tumor cell lines, including L5178Y, HeLa, and PC12 cells, with 7 being the most active (EC(50) against L5178Y cell line: 0.6 microg/mL).

The essential oils are well known for their antibacteric, antomycotic and insecticide effects. In this research the antomycotic activity of Rosemary essential oil has been tested in vitro versus different moulds, common contaminants of food and feed, as Aspergillus flavus, Aspergillus ochraceus, Aspergillus niger, Aspergillus ustus, Penicillium expansum, Penicillium aurantio-griseum, Fusarium moniliforme. The Rosemary essential oil tested (produced in Sardegna, Italy) shows a different efficacy against various moulds and his activity seems to be produced by borneol, α-pinene, eucalyptol, camphor and limonene. Aspergillus and Penicillium are the genus showing an important inhibition of their development in vitro. Rosemary essential oil for these natural properties can be used as antimycotic additive to extended shelf-life of many foodstuffs.

Aspergillus ustus is a relatively rare human pathogen causing invasive infections in immunocompromised hosts. In this study isolates originating from clinical and other sources have been examined using molecular, morphological, and physiological approaches to clarify their species assignment. Phylogenetic analysis of partial β-tubulin, calmodulin, actin, and intergenic transcribed spacer sequences indicated that none of the clinical isolates recognized previously as A. ustus belongs to this species. All but two of these isolates formed a well-defined clade related to A. pseudodeflectus based on sequence analysis of protein-coding regions. Morphological and physiological examination of these isolates indicated that they are able to grow above 37°C, in contrast with A. ustus isolates, and give a positive Ehrlich reaction, in contrast with related species including A. granulosus, A. ustus , and A. pseudodeflectus . These isolates are proposed as a new species, A. calidoustus . Antifungal susceptibility testing showed that this species has decreased susceptibilities to several antifungal drugs. The triazoles are inactive in vitro, including the new azole posaconazole.