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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.

Invasive aspergillosis (IA) is the most common invasive fungal infection following a hematopoietic cell transplant, with emerging cryptic species exhibiting resistance to commonly used antifungals such as azoles. These species have been increasingly found after the introduction of anti-mold prophylaxis. We report a case of a 56-year-old female with primary myelofibrosis whose allogeneic hematopoietic cell transplant was complicated by disseminated fungal infection (skin, lung) due to Aspergillus calidoustus, a cryptic specie. Treatment of Aspergillus species remains challenging as these cryptic species are usually resistant to azoles including voriconazole which is the first line of treatment of IA. Infection was successfully treated with surgical excision and combination antifungal therapy based on in vitro susceptibility and synergy testing. Therapy included isavuconazole, a drug that has been shown to be non-inferior to voriconazole in the treatment of invasive mold infections.

Background Aspergillus spp. of section Usti (A. ustus) represent a rare cause of invasive aspergillosis (IA). This multicenter study describes the epidemiology and outcome of A. ustus infections. Methods Patients with A. ustus isolated from any clinical specimen were retrospectively identified in 22 hospitals from 8 countries. When available, isolates were sent for species identification (BenA/CaM sequencing) and antifungal susceptibility testing. Additional cases were identified by review of the literature. Cases were classified as proven/probable IA or no infection, according to standard international criteria. Results Clinical report forms were obtained for 90 patients, of whom 27 had proven/probable IA. An additional 45 cases were identified from literature review for a total of 72 cases of proven/probable IA. Hematopoietic cell and solid-organ transplant recipients accounted for 47% and 33% cases, respectively. Only 8% patients were neutropenic at time of diagnosis. Ongoing antimold prophylaxis was present in 47% of cases. Pulmonary IA represented 67% of cases. Primary or secondary extrapulmonary sites of infection were observed in 46% of cases, with skin being affected in 28% of cases. Multiple antifungal drugs were used (consecutively or in combination) in 67% of cases. The 24-week mortality rate was 58%. A. calidoustus was the most frequent causal agent. Minimal inhibitory concentrations encompassing 90% isolates (MIC90) were 1, 8, >16, and 4 µg/mL for amphotericin B, voriconazole, posaconazole, and isavuconazole, respectively. Conclusions Aspergillus ustus IA mainly occurred in nonneutropenic transplant patients and was frequently associated with extrapulmonary sites of infection. Mortality rate was high and optimal antifungal therapy remains to be defined.

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) The contamination of agricultural lands by pesticides, their overuse and toxicity to many non-target organisms cause serious environmental problems. Therefore, the development of bioremediation methods for distinct active ingredients, such as pyrethroids, is primordial. In this work, the biodegradation of gamma-cyhalothrin (GCH) by fungi from the cave Gruta do Catão was investigated. Experiments were conducted with different fungi strains (Aspergillus ustus CBMAI 1894, Talaromyces brunneus CBMAI 1895 and Aspergillus sp. CBMAI 1926) in 2% Malt liquid medium with 300 mg L-1 of GCH in triplicate employing a validated method. All strains biodegraded this insecticide and the most efficient was Aspergillus ustus with a degradation rate of approximately 50% of the initial GCH concentration in 21 days of experiment. We also conducted an experimental design testing three variables: temperature (25-35 degrees), pH (5,5– 8,5) and pesticide concentration (50-550 mg L-1) being the temperature and pesticide concentration were those that most influenced the biodegradation. So, we concluded that cave fungi biodegraded GCH and can be employed in future studies for enzymatic characterization and bioremediation of contaminated environments. A contaminação de áreas agriculturáveis por pesticidas, seu uso intensivo e toxicidade para muitos organismos não alvo causam sérios problemas ambientais. Portanto, o desenvolvimento de metodologias de biorremediação para ingredientes ativos distintos, tais como piretróides, é primordial. Nesse trabalho, a biodegradação de gama cialotrina (GCH) por fungos da Gruta do Catão foi investigada. Os experimentos foram realizados com três diferentes linhagens fúngicas (Aspergillus ustus CBMAI 1894, Talaromyces brunneus CBMAI 1895 e Aspergillus sp. CBMAI 1926) em meio líquido malte 2% com 300 mg L-1 de GCH em experimentos em triplicata utilizando um método validado. Todas as linhagens biodegradaram esse inseticida e a espécie mais eficiente foi Aspergillus ustus com uma taxa de degradação de aproximadamente 50% da concentração inicial de GCH em 21 dias de experimento. Também conduzimos um planejamento experimental testando três variáveis: temperatura (25-35 graus), pH (5,5 - 8,5) e concentração de pesticida (50-550 mg L-1), sendo que a temperatura e a concentração influenciaram no experimento. Concluímos que fungos de caverna biodegradaram GCH e podem ser empregados em estudos futuros para caracterização enzimática e biorremediação de ambientes contaminados. CNPq: 144586/2017-7

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.

A co-cultivation study of two fungal strains showed that Aspergillus ustus could inhibit Aspergillus repens growth. The bioactive compound responsible for the observed activity was purified and identified as a sesterterpene, ophiobolin K. Ophiobolin K exhibited marked inhibition against both fungi and bacteria, especially A. repens, A. glaucus and gram-positive bacteria including Bacillus subtilis, Staphylococcus aureus, and Micrococcus luteus.

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.

Progesterone was biotransformed byAspergillus ustusin this paper. Two products were obtained and they were identified as 11α-hydroxyprogesterone and 11α,15β-dihydroxyprogesterone by NMR and MS. The fermentation process of biotransformtion had been studied with HPLC and 11α-hydroxyprogesterone was the main product.

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.

Heat inactivated Aspergillus ustus (Asp), silicon dioxide-nano-powder (N―Si), and silicon dioxide nano-powder-combined-heat inactivated Aspergillus ustus (N―Si―Asp) were used to study the biosorption of Cd(II) from aqueous solutions via batch equilibrium technique. Surface characterization and immobilization of the fungal cells on silicon dioxide-nano-powder were examined and confirmed by using FT-IR and ESM analysis. Cadmium biosorption processes were investigated under the effect of pH, contact time, sorbent dosage and initial metal concentration. The three examined sorbents were found to exhibit maximum μmol g-1 capacity values in pH 7.0. The maximum determined cadmium capacity by silicon dioxide-nano-powder (N―Si) (600 μmol g− 1) was found higher than that exhibited by the heat inactivated fungal cells (Asp) (400 μmol g− 1). However, (N―Si―Asp) exhibited the highest sorption cadmium capacity (1000 μmol g− 1) as a combined behavior of both silicon dioxide nano-powder and Aspergillus ustus units. Sorption equilibria were established in 20 min and their data were well described by both Langmuir and Freundlich models. The potential applications of these sorbents for biosorptive removal of Cd(II) from real samples contaminated with cadmium, were successfully accomplished via a micro-column under constant flow rate.

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.

Eight drimane sesquiterpenes (1-8), six isochromane derivatives (9-14), and three known compounds, daldinin B (15), 9alpha-hydroxy-6beta-[(2E,4E,6E)-octa-2,4,6-trienoyloxy]-5alpha-drim-7-en-11,12-olide (16), and pergillin (17), were isolated from the EtOAc extract of the marine-derived fungus Aspergillus ustus 094102. The structures of the new compounds were elucidated on the basis of spectroscopic analysis. The cytotoxic effects on A549 and HL-60 cell lines were evaluated by SRB and MTT methods. Ustusorane E (13) showed significant cytotoxicity against HL-60 cells with an IC50 value of 0.13 microM. Ustusolates C (6) and E (8) exhibited moderate cytotoxicity against A549 and HL-60 cells with IC50 values of 10.5 and 9.0 microM, respectively, and ustusolate A (4) showed weak cytotoxicity against HL-60 and A549 cells with IC50 values of 20.6 and 30.0 microM, respectively.

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.

This report describes the first case of cerebral aspergillosis in a heart transplantation patient caused by Aspergillus ustus and reviews 15 previously reported cases of invasive aspergillosis in immunocompromised hosts caused by this mold. The utility of molecular analysis for the identification of unusual fungal pathogens is also described. The refractory nature of A. ustus to treatment is similar to other Aspergillus species and treatment options are reviewed.

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.

Specimens of Suberites domuncula that had been cultured in aquaria for 4 weeks were analyzed for their associated fungi. A total of 81 fungal strains belonging to 20 different genera was isolated and identified by morphological and molecular methods. The most frequently isolated taxa were Cladosporium spp., Penicillium spp., Petriella sp., Phialophora spp. and Engyodontium album. Based on chromatographic and mass spectrometric analysis of fungal extracts, as well as on bioassay results, Aspergillus ustus, Penicillium sp., Petriella sp. and Scopulariopsis sp. were selected for in-depth analysis of their natural products. A total of 19 different fungal metabolites, including three new natural products, was isolated and structurally identified. A. ustus yielded two sesquiterpenes, a drimane derivative and deoxyuvidin, as well as a sesterterpene ophiobolin H. The drimane derivative had an ED50 value against L5178Y cells of 1.9 μg ml-1 in vitro. The crude extract of Petriella sp. was also strongly cytotoxic against the L5178Y cell line. The cyclic tetrapeptide WF-3161 was primarily responsible for the activity; the ED50 value was -1. It was identical to the known compound WF-3161 and had been previously isolated from Petriella guttulata. In addition to WF-3161, three further natural products were obtained and unequivocally identified as new derivatives of infectopyrone by one- and two-dimensional NMR spectroscopy and by mass spectroscopy. Of the new compounds, only dihydroinfectopyrone was active against L5178Y cells; the ED50 value was 0.2 μg ml-1. Penicillium sp. yielded the largest number of metabolites. Viridicatin, viridicatol, cyclopenin and cyclopenol suppressed larval growth of the polyphagous pest insect Spodoptera littoralis when incorporated into an artificial diet at an arbitrarily chosen concentration of 237 ppm. Viridicatol was the most active compound and had an ED50 value of ca. 50 ppm. Scopulariopsis sp. yielded three metabolites, including the known acetylcholinesterase inhibitors quinolactacin A1 and A2.

Meadow chernozemic soil was studied in an idle land-shelterbelt-cropland system. It was found that the long-term agricultural use of chernozems leads to a decrease in the total nitrogen content and to changes in the soil microbial community. The portion of phytopathogenic Fusarium, Aspergillus ustus, Penicillium daleae, and Penicillium rubrum microorganisms in the complex of soil fungi increases, and the phytosanitary state of the soil worsens. The use of biotests made it possible to estimate the role of particular microorganisms in the soil phytotoxicity.

The presence of moulds, potential producers of sterigmatocystine, was investigated in 20 samples of spices (cinnamon, caraway, marjoram and clove). The following five species were isolated from the samples: Aspergillus versicolor, Aspergillus sydowii, Aspergillus ustus, Emericella nidulans and Eurotium herbariorum. E. herbariorum was present in all the investigated spice samples, accounting for 7.6% of total mycopopulation in caraway and 42.9% in cloves. A. versicolor was detected in cinnamon, caraway and marjoram, accounting for 0.4% of total mycopopulation in caraway and 1.5% in cinnamon; A. ustus accounted for 0.1% of total mycopopulation in caraway and 0.4% in marjoram, while A. sydowii accounted for 18.9% of total mycopopulation in marjoram and 7.1% in clove. E. nidulans was a contaminant of caraway (0.01%) and marjoram (0.4%). .