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Aspergillus is a ubiquitous soil-dwelling organism found in organic debris, dust, compost, foods, spices, and rotted plants. The genus Aspergillus includes over 185 species. Around 20 species have so far been reported as causative agents of opportunistic infections in humans. Among these, Aspergillus fumigatus is the most commonly isolated species, followed by Aspergillus flavus and Aspergillus niger. Aspergillus oryzae, Aspergillus terreus, Aspergillus ustus, and Aspergillus versicolor are among the other species less commonly isolated as opportunistic pathogens (Rinaldi, 1983). Aspergillus spp. are well known to play a role in three different clinical settings in humans: (1) opportunistic infections, (2) allergic states, and (3) toxicoses. Immunosuppression is the major factor predisposing to development of opportunistic infections (Ho and Yuen, 2000). Aspergillus, like other lamentous fungi, is primarily acquired from an inanimate reservoir, usually by the inhalation of airborne spores. The organism grows best at 37°C, and the small spores (2-3 µm) are easily inhaled and deposited deep in the lungs, leading to a variety of clinical syndromes (Figure 14.1). These infections may present in a wide spectrum, varying from local involvement to dissemination and as a whole called aspergillosis. Among all lamentous fungi, Aspergillus is in general the most commonly isolated one in invasive infections. It is the second most commonly recovered fungus in opportunistic mycoses following Candida. Although these are distinct pulmonary entities, on rare occasions, one condition may change to another; for example, an aspergilloma may change to invasive pulmonary aspergillosis (IPA) (Tomee et al., 1995).

Secondary metabolites (SMs) produced by Aspergillus have been extensively studied for their crucial roles in human health, medicine and industrial production. However, the resulting information is almost exclusively derived from a few model organisms, including A. nidulans and A. fumigatus, but little is known about rare pathogens. In this study, we performed a genomics based discovery of SM biosynthetic gene clusters in Aspergillus ustus, a rare human pathogen. A total of 52 gene clusters were identified in the draft genome of A. ustus 3.3904, such as the sterigmatocystin biosynthesis pathway that was commonly found in Aspergillus species. In addition, several SM biosynthetic gene clusters were firstly identified in Aspergillus that were possibly acquired by horizontal gene transfer, including the vrt cluster that is responsible for viridicatumtoxin production. Comparative genomics revealed that A. ustus shared the largest number of SM biosynthetic gene clusters with A. nidulans, but much fewer with other Aspergilli like A. niger and A. oryzae. These findings would help to understand the diversity and evolution of SM biosynthesis pathways in genus Aspergillus, and we hope they will also promote the development of fungal identification methodology in clinic.

This is the first report of clustered Aspergillus ustus causing systemic disease in transplant patients., Aspergillus ustus is a mold that rarely infects humans; only 15 systemic cases have been reported. We report the first outbreak of invasive infection caused by A. ustus among hematopoietic stem cell transplant (HSCT) recipients. Six patients with infections were identified; 3 infections each occurred in both 2001 and 2003. Molecular typing by using randomly amplified polymorphic DNA (RAPD) and antifungal drug susceptibility testing were performed on clinical and environmental isolates recovered from our hospital from 1999 to 2003. The highest overall attack rate in HSCT patients was 1.6%. The overall death rate was 50%, and death occurred within 8 days after diagnostic culture collection. Clinical isolates exhibited decreased susceptibility to antifungal drugs, especially azoles. RAPD and phylogenetic analysis showed genetic similarity between isolates from different patients. Based on the clustering of cases in space and time and molecular data, common-source acquisition of this unusual drug-resistant species is possible.

The first case of postoperative endophthalmitis after cataract surgery caused by Aspergillus ustus, a species that has only rarely been implicated in human disease, is described. Six weeks after cataract surgery, a 67-year-old medically controlled diabetic patient presented with uveitis, mild ciliary injection and ocular discomfort. Anterior chamber paracentesis, vitreous tap and finally complete vitrectomy with removal of the capsular bag including the intraocular lens were performed and several sets of culture yielded A. ustus. Despite vigorous systemic (itraconazole and caspofungin) and intravitreal (amphotericin B and caspofungin) antifungal therapy, the endophthalmitis did not improve. The painful eye with marked inflammation was finally enucleated. In vitro susceptibility testing of the isolate showed that it appeared resistant to amphotericin B, caspofungin, itraconazole, voriconazole, and posaconazole, and susceptible to terbinafine.

Molecular approaches are now being developed to provide a more rapid and objective identification of fungi compared to traditional phenotypic methods. Ribosomal targets, especially the large-subunit RNA gene (D1-D2 region) and internal transcribed spacers 1 and 2 (ITS1 and ITS2 regions), have shown particular promise for the molecular identification of some fungi. We therefore conducted an assessment of these regions for the identification of 13 medically important Aspergillus species: Aspergillus candidus , Aspergillus ( Eurotium ) chevalieri , Aspergillus ( Fennellia ) flavipes , Aspergillus flavus , Aspergillus fumigatus , Aspergillus granulosus , Aspergillus ( Emericella ) nidulans , Aspergillus niger , Aspergillus restrictus , Aspergillus sydowii , Aspergillus terreus , Aspergillus ustus , and Aspergillus versicolor . The length of ribosomal regions could not be reliably used to differentiate among all Aspergillus species examined. DNA alignment and pairwise nucleotide comparisons demonstrated 91.9 to 99.6% interspecies sequence identities in the D1-D2 region, 57.4 to 98.1% in the ITS1 region, and 75.6 to 98.3% in the ITS2 region. Comparative analysis using GenBank reference data showed that 10 of the 13 species examined exhibited a ≤1-nucleotide divergence in the D1-D2 region from closely related but different species. In contrast, only 5 of the species examined exhibited a ≤1-nucleotide divergence from sibling species in their ITS1 or ITS2 sequences. Although the GenBank database currently lacks ITS sequence entries for some species, and major improvement in the quality and accuracy of GenBank entries is needed, current identification of medically important Aspergillus species using GenBank reference data seems more reliable using ITS query sequences than D1-D2 sequences, especially for the identification of closely related species.

Suppression of plant parasitic nematodes with nematode predators, parasites or antagonists is an eco-friendly approach than the toxic chemicals. In a study, soil borne fungi from the rhizosphere of major spice crops were collected from diverse cropping systems prevailing in three southern states of India. A series of in vitro studies were conducted using 73 freshly collected fungal isolates and 76 isolates obtained from other sources. Out of this 67 isolates were not parasitic on females of root-knot nematodes whereas 115 isolates, though colonized the egg masses, did not show any signs of parasitism on nematode eggs. Fifty-nine isolates showed 50-90% inhibition in egg hatch. Pochonia chlamydospora, Verticillium lecanii, Paecilomyces lilacinus, and few isolates of Trichoderma spp. showed >25% parasitism on root-knot nematode eggs. The most promising isolates in this study were one isolate each of Aspergillus (F.45), Fusarium (F.47), and Penicillium (F.59); three each isolates of Trichoderma (F.3, F.52, and F.60) and Pochonia (F.30 and Vc.3) Verticillium (Vl); and two isolates of fungi that could not be identified (F.28 and F.62). Parasitism by Aspergillus tamarii, Aspergillus ustus, Drechslera sp., Humicola sp., and Scopulariopsis sp. on root-knot nematode eggs or females, reported in the present study, are new reports.

We developed a PCR-based assay to differentiate medically important species of Aspergillus from one another and from other opportunistic molds and yeasts by employing universal, fungus-specific primers and DNA probes in an enzyme immunoassay format (PCR-EIA). Oligonucleotide probes, directed to the internal transcribed spacer 2 region of ribosomal DNA from Aspergillus flavus , Aspergillus fumigatus , Aspergillus nidulans , Aspergillus niger , Aspergillus terreus , Aspergillus ustus , and Aspergillus versicolor , differentiated 41 isolates (3 to 9 each of the respective species; P < 0.001) in a PCR-EIA detection matrix and gave no false-positive reactions with 33 species of Acremonium , Exophiala , Candida , Fusarium , Mucor , Paecilomyces , Penicillium , Rhizopus , Scedosporium , Sporothrix , or other aspergilli tested. A single DNA probe to detect all seven of the most medically important Aspergillus species ( A. flavus , A. fumigatus , A. nidulans , A. niger , A. terreus , A. ustus , and A. versicolor ) was also designed. Identification of Aspergillus species was accomplished within a single day by the PCR-EIA, and as little as 0.5 pg of fungal DNA could be detected by this system. In addition, fungal DNA extracted from tissues of experimentally infected rabbits was successfully amplified and identified using the PCR-EIA system. This method is simple, rapid, and sensitive for the identification of medically important Aspergillus species and for their differentiation from other opportunistic fungi.

An effective method was established for preparing the potent cell cycle inhibitor dehydrophenylahistin by a combination of chemical racemization of partially purified (±)-phenylahistin and enzymatic conversion of (−)-phenylahistin by the cell-free extract of Streptomyces albulus KO-23, an albonoursin-producing actinomycete. This method enables conversion of (+)-phenylahistin, which is present in the culture of Aspergillus ustus NSC-F038 and is not transformed by the Streptomyces enzyme, to dehydrophenylahistin.

Fourwing saltbush, Atriplex canescens (Pursh) Nutt., is an ecologically important range plant in arid south-western U.S.A. rangelands. Native populations of this chenopodiacious shrub are more extensively colonized by melanized dark septate fungi (DS) than by conventional mycorrhizal fungi. Seedling radicles of A. canescens are colonized at germination by a DS fungus identified as Aspergillus ustus that cannot be removed by heat or sterilization. The association of A. canescens with A. ustus was evaluated by comparing naturally colonized control seedlings receiving no P (0P) or adequate plant available P (AAP) receiving 30 p.p.m. supplied as KH2PO4 in the root zone to seedlings whose roots were separated from plant unavailable P (as rock phosphate (RP) or tricalcium phosphate (TCP)) by a barrier that only allowed access by the fungus. A. ustus penetrated A. canescens roots with hyaline septate hyphae, formed melanized runner hyphae at the root surface, and extended through the root exclusion barrier into RP and TCP. In these treatments A. ustus obtained plant carbon, increased shoot and root biomass, and phosphorus use efficiency. A. ustus grew well in culture on RP and TCP and internally colonized in vitro Ri-T DNA D. carota roots. The mutualistic association of DS fungi in arid ecosystems is discussed.

The rhizospheric soil microfungi from a native forest (undisturbed and disturbed) were studied using soil dilution plate and soil washing methods. Fungi were isolated using slightly acid and alkaline culture media. 54 taxa were isolated: 49 from undisturbed forest soil and 37 from disturbed forest soil. Acremonium sp., Aspergillus ustus, Coemansia pectinata, Doratomyces stemonitis, Fusarium solani, F. oxysporum, Gliocladium roseum, Humicola fusco-atra, Mortierella sp., Penicillium lilacinum, Trichoderma harzianum, and T. koningii, showed the highest frequency, in both, undisturbed and disturbed forests. In undisturbed soil forest the biodiversity index was 3.97 whereas in disturbed ones was 3.89., Instituto de Botánica "Dr. Carlos Spegazzini"

Aspergillus ustus is among the most ubiquitous soil species and has been implicated in human infections. The complete mitochondrial genome of A. ustus has been determined by high-throughput sequencing technology in this work. Our study revealed that the mitochondrial genome of A. ustus is 33,007 bp long, with AT content of 74.84%, which consists of a usual set of mitochondrial proteins and RNA genes, including large and small ribosomal RNAs, 15 proteins and 20 tRNA genes and contains two introns. Notably, it also contains three hypothetical proteins without obvious homology to any known proteins. All structural genes are located on one strand and are apparently transcribed in one direction. The complete mitochondrial genomes of A. ustus would be useful for future investigation of the genetics, evolution and clinical identification of Aspergillus species.

This chapter focuses on issues related to collection and isolation of deep-sea fungi, direct detection in deep-sea sediments, diversity and biomass, growth and physiology, adaptations, and their biotechnological applications. The presence of fungi in oceanic waters and the deep sea has been sporadically reported in the past. Their presence in shells collected from deep-sea waters at a depth of 4,610 m was the first report on deep-sea fungi. Immunofluorescence has been widely used to detect specific fungi in terrestrial and in a few marine substrates. The authors used this for detecting one of the commonly isolated fungi, Aspergillus terreus (isolate A 4634), from deep-sea sediments of the Central Indian Basin. The deep-sea fungi when grown under elevated pressure synthesized extracellular protease, albeit in very low quantities in comparison with that produced under 0.1 MPa. Aspergillus ustus (NIOCC20) isolated by the authors from deep-sea sediments produced cold-active alkaline serine protease, whereas A. ustus obtained from a terrestrial habitat did not. This might be due to differences in strains but may also indicate the adaptation that deep-sea fungi have undergone for their survival. Study of the cold shock or stress proteins or genes produced in response to hydrostatic pressure shock in fungi using proteomics and microarray technology will help to understand the response in eukaryotic organisms to pressure. New techniques for retrieval of sediment samples with in situ pressure, isolation, and culture of the vast diversity of organisms from the deep sea will open new vistas in deep-sea biology.

A colorimetric assay for antifungal susceptibility testing of Aspergillus species ( Aspergillus fumigatus , Aspergillus flavus , Aspergillus terreus , Aspergillus nidulans , and Aspergillus ustus ) is described based on the reduction of the tetrazolium salt 2,3-bis(2-methoxy-4-nitro-5-[(sulphenylamino)carbonyl]-2H-tetrazolium-hydroxide (XTT) in the presence of menadione as an electron-coupling agent. The combination of 200 μg of XTT/ml with 25 μM menadione resulted in a high production of formazan within 2 h of exposure, allowing the detection of hyphae formed by low inocula of 10 2 CFU/ml after 24 h of incubation. Under these settings, the formazan production correlated linearly with the fungal biomass and less-variable concentration effect curves for amphotericin B and itraconazole were obtained.

The mode of attachment of dimethylallyl pyrophosphate (DMAPP) in the biosynthesis of the indole alkaloids paraherquamide A, austamide, and brevianamide A has been studied. Feeding experiments on Penicillium fellutanum, Penicillium brevicompactum, and Aspergillus ustus using [13C2]-acetate showed isotopic scrambling of the geminal methyl groups originating from C-2 of the indole ring precursors in paraherquamide A, brevianamide A, and austamide biosynthesis. The labeling patterns suggest that the methyl groups of dimethylallyl pyrophosphate become equivalent during the biosyntheses; a non-face-selective SN‘ mechanism has been invoked to account for these observations.

Three novel isoquinoline alkaloids, TMC-120A, B and C (1–3) have been isolated from a fermentation broth of Aspergillus ustus (Bain.) Thom & Church TC 1118, a fungus isolated from rhizosphere of grass. Their structures were determined through extensive spectroscopic analyses and chemical studies. TMC-120s (1–3) are found to be the first furo[3,2-h]isoquinoline-type alkaloids isolated from natural sources.

A case of invasive pulmonary aspergillosis in an allogeneic bone marrow transplant recipient caused by Aspergillus ustus is presented. A. ustus was also recovered from the hospital environment, which may indicate that the infection was nosocomially acquired. A literature review revealed seven cases of invasive infections caused by A. ustus , and three of these were primarily cutaneous infections. In vitro susceptibility testing of 12 A. ustus isolates showed that amphotericin B and terbinafine had fungicidal activity and that itraconazole and voriconazole had fungistatic activity.

In this study, the ability to produce mycotoxins during growth on artificially infested building materials was investigated for Penicillium chrysogenum, Pen. polonicum, Pen. brevicompactum, Chaetomium spp., Aspergillus ustus, Asp. niger, Ulocladium spp., Alternaria spp., and Paecilomyces spp., all isolated from water-damaged building materials. Spores from the different isolates of the above mentioned species were inoculated on gypsum board with and without wallpaper and on chipboard with and without wallpaper. Fungal material was scraped off the materials, extracted, and analyzed using high performance liquid chromatography-diode array detection and thin layer chromatography. All six isolates of C. globosum produced the toxic chaetoglobosins A and C, at levels of up to 50 and 7 μg/cm2 respectively. The quantities of secondary metabolites produced by Penicillia were generally low, and no toxin production was detected from any of the five isolates of Pen. chrysogenum. Both isolates of Pen. polonicum produced 3-methoxy-viridicatin, verrucosidin, and verrucofortine. Two of five isolates of Pen. brevicompactum produced mycophenolic acid. From five out of six isolates of Alternaria spp., alternariol and alternariol monomethyl ether were detected. From Ulocladium spp., Paecilomyces spp., and Asp. ustus no known mycotoxins were detected, although the latter two are known mycotoxin producers. Asp. niger produced several naphtho-γ-pyrones and tetra-cyclic compounds. All investigated species, especially Asp. ustus and Asp. niger produced many unknown secondary metabolites on the building materials. Analyses of wallpaper and glass-fibre wallpaper naturally infested with Asp. versicolor revealed sterigmatocystin and 5-methoxysterigmatocystin. Analyses of naturally infested wallpaper showed that C. globosum produced the chaetoglobosins A and C, and Pen. chrysogenum produced the antibiotic meleagrin.

A fungus biosensor based on an oxygen electrode and Aspergillus ustus immobilized in poly(vinyl alcohol) was developed to determine tannic acid in a batch system. The optimum temperature was 30.0°C and the optimum pH was 7.0 in 0.10 M phosphate buffer. The regression equation of the calibration curve between 0.0250 and 1.025 mM was (x ln y ) (−0.72724− x 0.53 /54) + C ( x ln y ) (−0.21891− x 0.08 /2) = A + B/x ( r ≥ 0.99985). The sensor had a life of at least 2 weeks; it yielded a negligible response to the compounds commonly coexisting with vegetable tannins.

Two species of filamentous fungi, AspergiUus ustus (Bain.) Thom & Church and Graphium sp., were isolated from calcareous animal shells at depths of 860 m in the Arabian Sea and 965 m in the Bay of Bengal. Laboratory experiments showed germination of conidia, growth of hyphae and microconidiation in both the fungi at 100 bar pressure at 10 and 30°C suggesting barotolerance The fungi also secreted barotolerant protease under these conditions. Protease synthesised and secreted in cultures grown at 1 bar was also active under 100 bar pressure at 10 and 30°C. At 1O0C, the fungi showed better conidial germination, growth and protease secretion when subjected to 100 bar than when grown at 1 atmosphere. The results indicate barotolerance of terrestrial species of fungl and suggest that they might be active under deep-sea conditions.

(−)-Phenylahistin is a fungal diketopiperazine metabolite consisting of L -phenylalanine and isoprenylated dehydrohistidine, and it showed an inhibitory activity on the cell cycle progression of P388 cells in the G2/M phase.

Aspergillus ustus is an uncommon clinical species which is poorly susceptible to antifungals. We report two cases of A. ustus infections that occurred in allogeneic stem cell transplant recipients while they were receiving either voriconazole or caspofungin. Prolonged use of these new antifungal agents may increase the risk of the emergence of resistant organisms.

Primary cutaneous aspergillosis is a rare disease usually caused by Aspergillus fumigatus, Aspergillus flavus, Aspergillus terreus and Aspergillus ustus. It is usually seen in immunocompromised hosts, though some cases are also reported in immunocompetent hosts. We present a case of an immunocompetent farmer who presented with generalised nodules and plaques, mimicking erythema nodosum leprosum but turned out to be cutaneous aspergillosis caused by Aspergillus tamarii. The characteristic ascospores of Aspergillus species were found in skin lesions on fungus isolated in culture. The patient showed excellent response to antifungal therapy.

A series of novel drimane sesquiterpene esters (1-6) was isolated from fermentations of Aspergillus ustus var. pseudodeflectus and their structures elucidated by spectroscopic methods including the HMQC, HMBC and INADEQUATE NMR experiments. The major component of the fermentation, 1, was (2'E,4'E,6'E)-6-(1'-carboxy-2',4',6'-trien)-9-hydroxydrim-7-ene-11 ,12-olide. Compounds 1, 2, 3 and 5 exhibited endothelin receptor binding inhibitory activity against rabbit endothelin-A and rat endothelin-B receptors with IC50 values in the range 20-150 microM. These compounds had similar levels of activity in assays for binding to human endothelin A and endothelin B receptors. The isolation of 9,11-dihydroxy-6-oxodrim-7-ene, 7, a probable biosynthetic precursor to the drimane esters is also reported.

Using gellan gum as a substitute for agar-agar in a mycological medium and sequencing of the ITS 1 and 2 regions resulted in an accurate identification of Aspergillus fumigatus, Aspergillus nidulans, Aspergillus terreus and Aspergillus ustus within 24 h of subculture.

Moulds constitute the largest group of bacteria, prevailing both in the indoor and outdoor air. Approximately 200 000 species of moulds have been identified so far, where only a small group of around 200 may present a threat to human health. Fungi from Aspergillus species are among the moulds considered to be most pathogenic. They also constitute the group of most pathogenic moulds most frequently isolated from the environment. Over 250 types of this species are known; about 50 of them were precisely described before the year 2000 (Klich, 2009). Pathogenicity, due to their toxicity, was also documented in other species: Aspergillus fumigatus, Aspergillus flavus, Aspergillus ochraceus, Aspergillus niger, Aspergillus versicolor, Aspergillus parasiticus. Aspergillus nidulans, Aspergillus ustus, Aspergillus glaucus, Aspergillus clavatus, Aspergillus sydowii and Aspergillus terreus. Their taxonomic identification is still an open topic because of their morphological variability and ability to produce metabolites; new species which exhibit adverse health effects to humans are constantly being detected. Aspergillus lentulus is one of the recently detected species of considerable clinical importance; it reveals similarity to Aspergillus fumigatus, one of the most pathogenic fungus for humans (Balajee et al., 2005).Table 1 presents the classification of species of fungi pathogenic to humans from the Aspergillus species. The list has been created by the authors of “Atlas Grzybow Chorobotworczych Czlowieka" ("Atlas of Fungi Pathogenic to Humans") (Krzyściak et al., 2011).

The new compounds (I)-(V) and 14 known analogues are screened for their antitumor activity against P388, HL-60, K562, and BEL-7402 cell lines.

Fungi present in the rhizosphere and rhizoplane of Capsicum frutescens at different stages of plant growth were studied. The experimental soil was loamy sand in texture. The pH of the rhizosphere soil in the field ranged from 6.4 to 6.7 and in the pots, it ranged from 6.3 to 6.8. The fungi isolated were Aspergillus niger, Rhizopus stolonifer, Aspergillus flavus, Aspergillus candidus, Penicillium expansum, Fusarium oxysporum, Fusarium poae, Penicillium citrinum, Mucor racemosus, Mucor mucedo, Verticillium lateritium, Fusarium avenaceum, Trichophyton mentagrophyte, Fusarium verticilliodes, Aspergillus ustus, Saccharomyces cerevisiae and Aspegillus fumigatus . The most frequently isolated fungi were Rhizopus stolonifer, Aspergillus nigers and Mucor mucedo . The highest fungal number occurred on the 30th day of plant growth for pots and field. Key words : Fungi , rhizosphere, rhizoplane, plant

To deal with pathogens, plants have evolved sophisticated mechanisms including constitutive and induced defense mechanisms. Phytohormones play important roles in plant growth and development, as well as in the systemic response induced by beneficial and pathogen microorganisms. In this work, we identified an Aspergillus ustus isolate that promotes growth and induces developmental changes in Solanum tuberosum and Arabidopsis thaliana. A. ustus inoculation on A. thaliana and S. tuberosum roots induced an increase in shoot and root growth, and lateral root and root hair numbers. Assays performed on Arabidopsis lines to measure reporter gene expression of auxin-induced/ repressed or cell cycle controlled genes (DR5 and CycB1, respectively) showed enhanced GUS activity, when compared with mock-inoculated seedlings. To determine the contribution of phytohormone signaling pathways in the effect elicited by A. ustus, we evaluated the response of a collection of hormone mutants of Arabidopsis defective in auxin, ethylene, cytokinin, or abscisic acid signaling to the inoculation with this fungus. All mutant lines inoculated with A. ustus showed increased biomass production, suggesting that these genes are not required to respond to this fungus. Moreover, we demonstrated that A. ustus synthesizes auxins and gibberellins in liquid cultures. In addition, A. ustus induced systemic resistance against the necrotrophic fungus Botrytis cinerea and the hemibiotrophic bacterium Pseudomonas syringae DC3000, probably through the induction of the expression of salicylic acid, jasmonic acid/ethylene, and camalexin defense-related genes in Arabidopsis.

A Penicillium polonicum, an Aspergillus ustus and a Periconia britannica strain were isolated from water-damaged environments and the production of microbial volatile organic compounds (MVOCs) was investigated by means of headspace solid-phase microextraction followed by GC-MS analysis. The most important MVOCs produced were 2-methylisoborneol, geosmin and daucane-type sesquiterpenes for P. polonicum, 1-octen-3-ol, 3-octanone, germacrene D, ?-cadinene and other sesquiterpenes for A. ustus and the volatile mycotoxin precursor aristolochene together with valencene, ?-selinene and ?-selinene for P. britannica. Different growth conditions (substrate, temperature, relative humidity) were selected, resembling indoor parameters, to investigate their influence on fungal metabolism in relation with the sick building syndrome and the results were compared with two other fungal strains previously analyzed under the same conditions. In general, the range of MVOCs and the emitted quantities were larger on malt extract agar than on wallpaper and plasterboard, but, overall, the main MVOC profile was conserved also on the two building materials tested. The influence of temperature and relative humidity on growth and metabolism is different for different fungal species, and two main patterns of behavior could be distinguished. Results show that, even at suboptimal conditions for growth, production of fungal volatiles can be significant.

Recent studies have changed the taxonomy of Aspergillus section Usti, and a novel species, Aspergillus calidoustus, has been erected. It was also demonstrated that clinical isolates previously identified as A. ustus actually belong to the emerging pathogen A. calidoustus. Aspergillus ustus were frequently isolated from Norwegian water systems, and due to the taxonomical progress, these waterborne strains could be identified more precisely. A MLST study including ITS, calmodulin, β-tubulin and actin sequences was conducted on 32 strains previously identified as A. ustus. All strains were identified as A. calidoustus, which was verified by physiological, biochemical and phylogenetic analyses. This is the first report of that A. calidoustus is able to colonize water distribution systems. In respect to the potential role of water systems as a source of nosocomial infections in patients with immunodeficiency, attention should be given to water systems in hospitals and other healthcare units, especially the heated-water installations.

In continuation of research on secondary metabolites of marine fungi and actinobacteria [1, 2], hexane fractions and fractions of free fatty acids and the sterol fraction of actinobacterium were obtained and analyzed from cultures of marine isolates of the fungus Aspergillus ustus KMM 4642 and KMM 4664 isolated from sediment (Okhotsk Shelf, Sakhalin Island, 26.5 m depth) and the actinobacterium Nocardiopsis umidischolae KMM 7036 isolated from Mycale sp. of sponge (Deryugin Basin, Okhotsk Sea). Strain KMM 4642 was cultivated in malt-agar medium prepared with sea water for 14 d [3] and on rice medium prepared with seawater for 21 d [4]. Strain KMM 4664 was cultivated only on rice medium prepared with seawater. Actinobacterium was cultivated in a special medium containing peptone (5.0 g/L), meat extract (3.0), starch (20.0), agar (16.0) and seawater at pH 7.5 and 23°C for 23 d. Cultures were extracted with EtOAc. The extracts were evaporated to dryness. The resulting residues were dissolved in EtOH (10%) and extracted successively with hexane, EtOAc, and BuOH. The hexane fractions were evaporated at reduced pressure and analyzed by GC-MS. The data were compared with the mass spectrometric fragmentation of standards using the NIST98 database. The hexane fraction of A. ustus KMM 4642 consisted of 96.88% diethylhexylphthalate. The remainder consisted of fatty acid ethyl esters and pentadecane. The hexane fraction of A. ustus KMM 4664 contained fatty acid ethyl esters, squalene, octadecane, and sterols such as 24-cholest-4-en-3-one, 24-ethylcholest-7-en-3 -ol, 24-ethylcholesta-3,5-diene, and 24-ethylcholest-5-en-3 -ol at concentrations of 1–2% of the studied mixture. These results differed from those obtained earlier for marine isolates of A. ustus [1]. The hexane fraction of actinobacterium N. umidischolae contained hydrocarbons at concentrations of 1.5–3% that included linear C15, C16, C17, C18, C20, and C22; C18, C19, C20, C22, and C24 with an iso-carbon chain; C18, C19, and C22 with a terminal double bond; and dibutylphthalate (5%) and diethylhexylphthalate (45%). The EtOAc fraction of each culture was chromatographed over a column of silica gel using a hexane–EtOAc gradient (100:0 90:10) to afford fractions of free fatty acids. The obtained total acids were analyzed as the methyl esters (methylated by diazomethane in Et2O) and pyrrolides [5] using GC-MS. Derivatives were identified by comparing their mass spectra with those of standards using the NIST98 database. The composition and content of A. ustus acid esters are given below (mass%)

We report a case of primary cutaneous infection by the emerging fungus Aspergillus ustus in an immunosuppressed patient after a domestic accident. The patient failed to respond to itraconazole and died before receiving a new treatment with amphotericin B. There have been eight other cases reported since 1973, and only two patients survived the infection. In vitro susceptibility testing of seven antifungal drugs showed that terbinafine and the new azole derivative UR-9825 were the most active against this fungus.

The structure, stereochemistry, solution conformation and nematocidal activity of Ophiobolin K (1a) and 6-epiophiobolin K (1b) isolated from Aspergillus ustus has been described. The molecular structure of these compounds were deduced from extensive use of 2D NMR spectroscopy. The stereochemistry and solution conformation was determined in CDCl3 by the application of extensive 1H-1H NOE difference spectroscopy. Significant difference in the conformation of A/B cis 1a and its A/B trans isomer 1b was observed including the folding of the side chain. Ophiobolin K showed a unique boat like conformation whereas the B ring of 6- epiophiobolin K must exist in at least two different conformations. Ophiobolin K exhibited nematocidal activity (ED50 10 μg/mL) against the free-living nematode Caenorhabditis elegans , but the A/B trans isomer 1b was found to be inactive.

(−)-Phenylahistin is a fungal diketopiperazine metabolite consisting of L -phenylalanine and isoprenylated dehydrohistidine, and it showed an inhibitory activity on the cell cycle progression of P388 cells in the G2/M phase.

Three novel isoquinoline alkaloids, TMC-120A, B and C (1–3) have been isolated from a fermentation broth of Aspergillus ustus (Bain.) Thom & Church TC 1118, a fungus isolated from rhizosphere of grass. Their structures were determined through extensive spectroscopic analyses and chemical studies. TMC-120s (1–3) are found to be the first furo[3,2-h]isoquinoline-type alkaloids isolated from natural sources.

Summary 14 species belonging to 11 genera were identified from biodeteriorated paint film. The most fungi were Aspergillus ustus, A. niger, Chaetomium globosum, Cochliobolus spicifer and Curvularia lunata . All isolated fungi have endoglucanase (EC 3.2.1.4) and cellobiohydrolase (EC 3.2.1.91) activity. 10 −2 M of Ca +2 , Hg +2 and Cu +2 inhibited these activities.

Endo-polygalacturonase from Aspergillus ustus when immobilized on to modified silica gel retained 28% of its original activity. The immobilized enzyme could be re-used through 10 cycles of reaction with almost 90% retention of its original activity. It had increased thermostability over its soluble form: the half-life of the soluble enzyme at 40 °C was less than 10 h whereas the immobilized enzyme retained 82% of its activity after 10 h at 40 °C. Similarly, at 50 °C the half-life of the soluble enzyme was 30 min whereas that of the immobilized enzyme was 5 h.

Lung infections with Nocardia and Aspergillus spp in lung transplant recipients (LTRs) create diagnostic and therapeutic challenges. The present case illustrates the difficulties in identifying these pathogens in LTRs. A high degree of clinical suspicion and aggressive early management are required to ensure good outcomes. Although prospective data on treating these conditions are scarce, the empiric use of combination broad-spectrum anti-microbials initially seems prudent.

Introduction The purpose of this study was to research the presence of filamentous fungi in the root canals of teeth with pulp necrosis and periapical lesions and to perform the taxonomic study of the isolates. Methods Samples collected from 60 root canals with sterile absorbent paper points were inoculated in Sabouraud dextrose agar supplemented with chloramphenicol. The material was cultured from 7–14 days at room temperature to observe mycelial growth. Samples with growth were seeded in media specific for microculture (CYA agar, malt extract agar, or potato dextrose agar). Taxonomy was evaluated by macroscopic examination and optical microscopy. Results Filamentous fungi were isolated in situ from 17 of 60 samples (28.3%). Considering the positive root canal isolates, the genus Aspergillus was isolated from 7 samples (41%); 4 species were identified: Aspergillus ustus, A. granulosus, A. niger, and A. sydowii. Emericella quadriluniata , sexual form of Aspergillus , was isolated from 1 sample. Penicillium species ( Penicillium implicatum, P. micsynvisk, P. lividum, and P. citrionigrum ) were isolated from 4 samples (24%). Fusarium species ( Fusarium moniliforme and F. melanochorum ) were isolated from 2 samples (12%). The species Aureobasidium pullulans , Exophiala jeanselmei , Eurotium amstelodame, and Cladosporium sphaerospermum were isolated from 1 sample each. Conclusions The root canals of teeth with pulp necrosis and periapical lesions might present positive culture for filamentous fungi.

Made available in DSpace on 2016-09-23T14:00:15Z (GMT). No. of bitstreams: 1 relatorio_finalrebeca_pibic.pdf: 693157 bytes, checksum: 90d9785671ec90f6bdd5315f7a59ee08 (MD5) Previous issue date: 2009-07-31 CNPQ As crescentes preocupações com as questões ambientais, com a qualidade dos produtos e com o consumo de energia têm incentivado a busca de tecnologias limpas , sofisticadas e baratas. Neste contexto, a tecnologia enzimática tem ganhado espaço, isoladas ou juntamente com células íntegras, as enzimas são usadas como biocatalizadores em processos industriais bem como na despoluição ambiental. As enzimas têm elevado poder catalítico, sendo muitas vezes superiores aos catalisadores inorgânicos, além de alta especificidade por seus substratos, sendo assim não geradores de produtos indesejáveis, e podem atuar sob condições suaves de temperatura, pH e pressão, resultando em significante conservação de água, equipamentos e energia, para benefícios tanto da indústria, quantos do meio ambiente. Elas são facilmente degradadas e assimiladas pela cadeia alimentar, não contaminam lençóis freáticos, não são carcinogênicas, portanto, não poluem o ambiente.A β-glucosidase, desempenha um papel crucial na degradação enzimática da celulose, quebrando a celobiose até monômeros de glicose, tem um grande potencial de aplicação no processamento de bebidas, como vinho e suco de laranja e em outros produtos alimentícios. Esta enzima é capaz de transformar antocianinas, pigmentos presentes em muitos vegetais, em antocianidinas e açúcares. A aglicona resultante é menos colorida e solúvel que as antocianinas, assim tende a precipitar, tornando-se um composto mais fácil de ser removido. A enzima, quando usada no processamento de sucos de laranja, promove a hidrólise das antocianinas e a remoção das agliconas insolúveis durante a filtração, prevenindo a descoloração dos sucos causada pelo aquecimento durante a pasteurização, devido a presença de antocianinas. Essa enzima também pode ser usada para conseguir a cor desejada em vinhos rose. Este trabalho visa a purificação e caracterização de enzima β-glicosidase pelo fungo Aspergillus ustus, o qual será seqüência do trabalho PIBIC 2007, que foi desenvolvido um estudo de otimização da produção e caracterização do complexo celulolítico excretado pelo referido fungo

This is a case report of a 61-year-old cardiac transplant patient who developed a disseminated infection involving the upper extremity with a rare fungus known as Aspergillus ustus. The patient was successfully treated with aggressive serial debridements, antifungal medications, and reduction of immunosuppression. With these interventions, the patient avoided amputation despite the aggressive nature of this infection.

As part of studies of metabolites from fungi common in the built environment in Canadian homes, we investigated metabolites from strains of three Eurotium species, namely E. herbariorum, E. amstelodami, and E. rubrum as well as a number of isolates provisionally identified as Aspergillus ustus. The latter have been recently assigned as the new species A. insuetus and A. calidoustus. E. amstelodami produced neoechinulin A and neoechinulin B, epiheveadride, flavoglaucin, auroglaucin, and isotetrahydroauroglaucin as major metabolites. Minor metabolites included echinulin, preechinulin and neoechinulin E. E. rubrum produced all of these metabolites, but epiheveadride was detected as a minor metabolite. E. herbariorum produced cladosporin as a major metabolite, in addition to those found in E. amstelodami. This species also produced questin and neoechinulin E as minor metabolites. This is the first report of epiheveadride occurring as a natural product, and the first nonadride isolated from Eurotium species. Unlike strains from mainly infection-related samples, largely from Europe, neither ophiobolins G and H nor austins were detected in the Canadian strains of A. insuetus and A. calidoustus tested, all of which had been reported from the latter species. TMC-120 A, B, C and a sesquiterpene drimane are reported with certainty for the first time from indoor isolates, as well as two novel related methyl isoquinoline alkaloids.

The structure, stereochemistry, solution conformation and nematocidal activity of Ophiobolin K (1a) and 6-epiophiobolin K (1b) isolated from Aspergillus ustus has been described. The molecular structure of these compounds were deduced from extensive use of 2D NMR spectroscopy. The stereochemistry and solution conformation was determined in CDCl3 by the application of extensive 1H-1H NOE difference spectroscopy. Significant difference in the conformation of A/B cis 1a and its A/B trans isomer 1b was observed including the folding of the side chain. Ophiobolin K showed a unique boat like conformation whereas the B ring of 6- epiophiobolin K must exist in at least two different conformations. Ophiobolin K exhibited nematocidal activity (ED50 10 μg/mL) against the free-living nematode Caenorhabditis elegans , but the A/B trans isomer 1b was found to be inactive.

Aspergillus ustus infections are associated with a high mortality in immunocompromised hosts, and the mold has decreased susceptibility to most antifungal drugs, especially azoles. We report primary cutaneous A. ustus infection in a patient who failed itraconazole therapy and was switched to voriconazole (VRC). During VRC therapy, the MICs of VRC, amphotericin B (AMB), caspofungin (CFG), and terbinafine (TBF) were 4, 2, 64, and 0.13 μg/mL, respectively. Because the MIC to VRC was high, TBF was added to VRC for synergy based on anecdotal data from other mycoses. After treatment with VRC and TBF for 5 months, MICs of VRC, AMB, CFG, and TBF of A. ustus were 8, 1, 64, and 4 μg/mL respectively. Although the MICs of VRC and TBF increased during antifungal therapy, the patient responded well to the combination antifungal therapy with surgical debridement. With a successful outcome despite high MICs and with limited therapeutic options currently available, we investigated the in vitro activity of posaconazole (PCZ) and VRC individually and in combination with AMB, CFG, or TBF using the fractional inhibitory concentration index (FICI) method. Combination of VRC with TBF showed synergistic activity (FICI = 0.5). Therefore, combination of VRC and TBF with surgical debridement, when appropriate, may be a viable treatment option for refractory A. ustus infections.

Fungal pellets of Aspergillus niger 405, Aspergillus ustus 326, and Stachybotrys sp. 1103 were used for the removal of humic substances from aqueous solutions. Batchwise biosorption, carried out at pH 6 and 25 degrees C, was monitored spectrophotometrically and the process described with Freundlich's model. Calculated sorption coefficients K(f) and n showed that A. niger exhibited the highest efficiency. A good match between the model and experimental data and a high correlation coefficient (R2) pointed out to judicious choice of the mechanism for removal of humic substances from the reaction medium. The sorption rate constants (k) for A. ustus and Stachybotrys sp. were almost equal, however higher than that for A. niger. Comparison of test results with the simulated ones demonstrated the applicability of the designed kinetic model for removal of humic substances from natural water by biosorption with fungal pellets. Different morphological structure of the examined fungal pellets showed that faster sorption does not imply the most efficient removal of humic substances. Desorption of humic substances from fungal pellets was complete, rapid, and yielded uniform results.