Culture Of Aspergillus Fumigatus Research Articles

Effect of ozone gas on cultures of Candida albicans and Aspergillus fumigatus: evaluation of two ozonation devices.

Candida albicans and Aspergillus fumigatus are two important agents of Healthcare-associated infections. This study aimed to evaluate the antifungal activity of ozone (O3) gas produced by two commercial devices against cultures of these two species. Sterile plastic plates were inoculated with C. albicans and A. fumigatus and placed on a countertop at three distances (30 cm, 1 m, and 2 m) and three positions in relation to the wall (near, middle, and away), considering the source of O3. Plates were exposed to O3 for one hour and incubated. After incubation, the counting of colony-forming units was performed. As a control, an inoculated plate was incubated, without being exposed to O3. Tests were carried out with two different devices (namely, Mod.I and Mod.II), with the air conditioner on and off, in triplicate. Both devices showed antifungal activity. Mod. I presented better results, due to a higher flow rate. The best activity was on plates at 30 cm, middle position. Contrarily, on plates at 2m, near the wall, the inhibition activity was lower. The best results were obtained with the air conditioner off. Candida albicans was more sensitive to O3 than A. fumigatus. This method of decontamination by O3 gas shows potential due to its fast and easy execution. The establishment of new protocols for hygiene and hospital disinfection using this approach should be considered, which may reduce environmental contamination by fungi and, consequently, the burden of fungal infections.

Aspergillus fumigatus secretes a protease(s) that displays in silico binding affinity towards the SARS-CoV-2 spike protein and mediates SARS-CoV-2 pseudovirion entry into HEK-293T cells

BackgroundThe novel coronavirus disease of 2019 (COVID-19) is an infectious disease caused by severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2). Data from the COVID-19 clinical control case studies showed that this disease could also manifest in patients with underlying microbial infections such as aspergillosis. The current study aimed to determine if the Aspergillus (A.) fumigatus culture media (i.e., supernatant) possessed protease activity that was sufficient to activate the SARS-CoV-2 spike protein.MethodsThe supernatant was first analysed for protease activity. Thereafter, it was assessed to determine if it possessed proteolytic activity to cleave a fluorogenic mimetic peptide of the SARS-CoV-2 spike protein that contained the S1/S2 site and a full-length spike protein contained in a SARS-CoV-2 pseudovirion. To complement this, a computer-based tool, HADDOCK, was used to predict if A. fumigatus alkaline protease 1 could bind to the SARS-CoV-2 spike protein.ResultsWe show that the supernatant possessed proteolytic activity, and analyses of the molecular docking parameters revealed that A. fumigatus alkaline protease 1 could bind to the spike protein. To confirm the in silico data, it was imperative to provide experimental evidence for enzymatic activity. Here, it was noted that the A. fumigatus supernatant cleaved the mimetic peptide as well as transduced the HEK-293T cells with SARS-CoV-2 pseudovirions.ConclusionThese results suggest that A. fumigatus secretes a protease(s) that activates the SARS-CoV-2 spike protein. Importantly, should these two infectious agents co-occur, there is the potential for A. fumigatus to activate the SARS-CoV-2 spike protein, thus aggravating COVID-19 development.

Determination of fungicidal effect of disinfectant ‘SanDezVet’ on sanitary significant test cultures of mold micromycetes of the genus Aspergillus Mich.

Studying the properties of new disinfectants is a crucial aspect of disinfectology research. During the investigation of the fungicidal properties of the ‘SanDezVet’ disinfectant, it was discovered that the recommended concentrations of 0.1% and 0.5%, at a temperature of 20.0 ± 0.5 °C and an exposure time of 60, 120, and 180 minutes, do not inhibit the growth of Aspergillus fumigatus, Aspergillus flavus, and Aspergillus niger test cultures. This conclusion was reached as continuous growth of micromycetes was observed in all drug dilutions. The disinfectant ‘SanDezVet’ at a concentration of 3.0% displayed fungistatic properties by significantly delaying the growth of experimental test cultures. A 5.0% solution of the disinfectant resulted in a complete delay in growth of museum strains of Aspergillus fumigatus, Aspergillus flavus, and Aspergillus niger. This indicates fungicidal properties when compared to the positive control. Based on the obtained results, it was observed that the optimal exposure times for disinfection measures with ‘SanDezVet’ are 60 and 120 minutes. ‘SanDezVet’ can be effectively used for fungal infections at different veterinary facilities

Induction of hydrolytic enzyme production and antibiosis via a culture of dual fungal species isolated from soil rich with the residues of woody plants in Saudi Arabia

Inducing hydrolytic enzymes production and antibiosis is an attractive process for industrial applications. The approach can be used to repress pathogenic microorganisms. Using a dual culture of Aspergillus fumigatus and Aspergillus flavus, the activities of cellulase, polygalacturonase, chitinase, β-1,3-glucanases, protease, xylanase, and β-glucosidase were 1.78, 3.87, 2.98, 2.79, 6.91, 2.89 U, and 1.43 U∙mg-1 of protein. Meanwhile, the activities were 0.87, 2.78, 0.58, 1.69, 4.45, 2.06, and 0.89 U.mg-1 of protein for A. flavus alone and 0.98, 2.98, 0.87, 1.89, 4.98, 2.58, and 0.91 U.mg-1 of protein for A. fumigatus alone. The cellulase, polygalacturonase, and chitinase activities were studied at different temperatures; 40 °C and 50 °C, which were better temperatures than 20 °C in terms of enzymes activity for A. fumigatus and the dual culture compared to A. flavus. The highest antimicromial activity was observed using the dual fungal culture, where the inhibition zones were 3.13, 3.47, 2.27, 1.77, 1.03, and 2 mm compared with A. fumigatus alone or A. flavus alone, against Staphylococcus aureus, Escherichia coli, Bacillus subtilis, Aspergillus fumigatus, Mucor circinelloides, and Fusarium moniliforme, respectively.

Invasive pulmonary aspergillosis in the COVID-19 era: An expected new entity.

ObjectivesInformation on the recently COVID‐19‐associated pulmonary aspergillosis (CAPA) entity is scarce. We describe eight CAPA patients, compare them to colonised ICU patients with coronavirus disease 2019 (COVID‐19), and review the published literature from Western countries.MethodsProspective study (March to May, 2020) that included all COVID‐19 patients admitted to a tertiary hospital. Modified AspICU and European Organization for Research and Treatment of Cancer/Mycoses Study Group (EORTC/MSG) criteria were used.ResultsCOVID‐19‐associated pulmonary aspergillosis was diagnosed in eight patients (3.3% of 239 ICU patients), mostly affected non‐immunocompromised patients (75%) with severe acute respiratory distress syndrome (ARDS) receiving corticosteroids. Diagnosis was established after a median of 15 days under mechanical ventilation. Bronchoalveolar lavage was performed in two patients with positive Aspergillus fumigatus cultures and galactomannan (GM) index. Serum GM was positive in 4/8 (50%). Thoracic CT scan findings fulfilled EORTC/MSG criteria in one case. Isavuconazole was used in 4/8 cases. CAPA‐related mortality was 100% (8/8). Compared with colonised patients, CAPA subjects were administered tocilizumab more often (100% vs. 40%, p = .04), underwent longer courses of antibacterial therapy (13 vs. 5 days, p = .008), and had a higher all‐cause mortality (100% vs. 40%, p = .04). We reviewed 96 similar cases from recent publications: 59 probable CAPA (also putative according modified AspICU), 56 putative cases and 13 colonisations according AspICU algorithm; according EORTC/MSG six proven and two probable. Overall, mortality in the reviewed series was 56.3%.ConclusionsCOVID‐19‐associated pulmonary aspergillosis must be considered a serious and potentially life‐threatening complication in patients with severe COVID‐19 receiving immunosuppressive treatment.

A Biosorption study of Lead by Aspergillus Fumigatus

Heavy metals liberated by various industrial and agricultural processes are the major pollutants in soil, agricultural, marine, and industrial and even treated waste waters. Lead is greatly used in many industrial applications such as a storage battery manufacturing, printing, fuels, photographic materials, pigments and explosive manufacturing Objective: To assess the ability of Aspergillus fumigatus for removal of metal contamination such as lead by biosorptionMethods: The pure culture of the Aspergillus fumigatus was used for biosorption. The initial samples were cultured on the bread as the fungal spores were given suitable environmental conditions i.e., temperature, moisture, pH etc. For obtaining the pure culture of the Aspergillus fumigatus culture media was prepared. The spores collected were then allowed to grow on a specific culture media in a sterile Petri plates under aseptic conditions. Inoculation will be carried out by using 20 loops of fungal spores spread on the culture medium. After inoculation these plates were incubated at 28°C for few days and the colonies of the fungal strain becomes visible after 48 hours. The prepared agar is poured in the sterilized Petri plates were allowed to cool and solidify. As a result of this centrifugation, the spores and agar were separated. After 12 days of incubation, one fourth portion of the agar plate was cut and placed into the falcon tubes containing 10 ml distilled water Results: The results revealed that the biosorption recovery rate is maximum at 120 mg\g of dry mass i.e. 3.4%. The above estimation shows that the percentage of biosorption is directly proportional to the biomass concentration.

P147 Longitudinal effects of ivacaftor on Aspergillus fumigatus serology and culture in eligible adults with cystic fibrosis

P147 Longitudinal effects of ivacaftor on Aspergillus fumigatus serology and culture in eligible adults with cystic fibrosis

Pro-Inflammatory Responses in Human Bronchial Epithelial Cells Induced by Spores and Hyphal Fragments of Common Damp Indoor Molds.

Damp indoor environments contaminated with different mold species may contribute to the development and exacerbation of respiratory illnesses. Human bronchial epithelial BEAS-2B cells were exposed to X-ray treated spores and hyphal fragments from pure cultures of Aspergillus fumigatus, Penicillum chrysogenum, Aspergillus versicolor and Stachybotrys chartarum. Hyphal fragments of A. fumigatus and P. chrysogenum induced expression and release of the pro-inflammatory cytokine interleukin (IL)-6 and the chemokine IL-8, while none of the other hyphal preparations had effects. Hyphal fragments from A. fumigatus and P. chrysogenum also increased the expression of IL-1α, IL-1β and tumor necrosis factor (TNF)-α, but these cytokines were not released. X-ray treated spores had little or no inflammatory potential. Attenuating Toll-like receptor (TLR)-2 by blocking antibodies strongly reduced the A. fumigatus and P. chrysogenum hyphae-induced IL-6 and IL-8 release, whereas TLR4 antagonist treatment was without effects. Untreated A. fumigatus spores formed hyphae and triggered expression of pro-inflammatory genes with similarities to the effects of hyphal fragments. In conclusion, while X-ray treated spores induced no pro-inflammatory responses, hyphal fragments of A. fumigatus and P. chrysogenum enhanced a TLR2-dependent expression and release of IL-6 and IL-8.

The antifungal activity of extracts of Osmundea pinnatifida, an edible seaweed, indicates its usage as a safe environmental fungicide or as a food additive preventing post-harvest fungal food contamination.

In the present work, we explored the antifungal activity of the wild edible seaweed Osmundea pinnatifida (Rhodophyta) collected from the Portuguese coast, which is used as a food seasoning in Scotland, Ireland and Portugal. We performed a sequential extraction of the seaweed components with methanol, dichloromethane and n-hexane. These extracts showed an antifungal activity against Alternaria infectoria and Aspergillus fumigatus. The n-hexane fraction of the seaweed inhibited the sporulation of Alternaria infectoria at 30 μg mL-1 and induced a statistically significant (P < 0.001) decrease in β-glucan content. Furthermore, liquid cultures of Aspergillus fumigatus supplemented with 10 μg mL-1 of the n-hexane fraction showed abnormal conidiophores, completely devoid of phialides and conidia associated with a decrease of 18.3% in the chitin content (P < 0.01). The n-hexane fraction analysis by GC-MS revealed that it includes palmitic acid (29.6%), phytol isomer 1 (12.8%), oleic acid (9.6%), stearic acid (6.2%) and d-(-)-tagatofuranose (4.1%), among other compounds present at lower concentrations. The present study reveals Osmundea pinnatifida as a promising source of biologically active compounds inhibiting fungal growth and conidiation, the main dispersal mechanism of filamentous fungi as Aspergillus fumigatus and Alternaria alternata, revealing its utility both as an environmental fungicide against fungal diseases and as a food preservative against fungal post-harvest food contamination.

Microbial volatile communication in human organotypic lung models

We inhale respiratory pathogens continuously, and the subsequent signaling events between host and microbe are complex, ultimately resulting in clearance of the microbe, stable colonization of the host, or active disease. Traditional in vitro methods are ill-equipped to study these critical events in the context of the lung microenvironment. Here we introduce a microscale organotypic model of the human bronchiole for studying pulmonary infection. By leveraging microscale techniques, the model is designed to approximate the structure of the human bronchiole, containing airway, vascular, and extracellular matrix compartments. To complement direct infection of the organotypic bronchiole, we present a clickable extension that facilitates volatile compound communication between microbial populations and the host model. Using Aspergillus fumigatus, a respiratory pathogen, we characterize the inflammatory response of the organotypic bronchiole to infection. Finally, we demonstrate multikingdom, volatile-mediated communication between the organotypic bronchiole and cultures of Aspergillus fumigatus and Pseudomonas aeruginosa.

Sputum Inflammatory Mediators Are Increased in Aspergillus fumigatus Culture-Positive Asthmatics.

Aspergillus fumigatus sensitization and culture in asthma are associated with disease severity and lung function impairment, but their relationship with airway inflammation is poorly understood. We investigated the profile of 24 sputum inflammatory mediators in A. fumigatus culture-positive or-negative moderate-to-severe asthmatics. Fifty-two subjects were recruited from a single center. A. fumigatus was cultured from 19 asthmatics. Asthma control, symptom score, lung function, and sputum cell count were not significantly different between the asthmatics with and without a positive A. fumigatus culture. All of the sputum mediators were numerically increased in subjects with a positive versus negative sputum A. fumigatus culture. Sputum TNF-R2 was significantly elevated (P=0.03) and the mediator that best distinguished A. fumigatus culture-positive from culture-negative subjects (receiver-operator characteristic area under the curve 0.66 [95% CI: 0.51 to 0.82, P=0.045]). A. fumigates-positive culture in moderate-to-severe asthma is associated with increased inflammatory sputum mediators.

Hyperbranched polyester poly(3-diethylaminepropionate)s and their copper(II) complexes: Synthesis, characterization and biological investigation

New polydentate ligands based on hyperbranched second and third generation polyesters containing terminal (3-diethylamino)propionate fragments and their polynuclear copper(II) complexes were synthesized and characterized by spectroscopic methods. The acid-base properties spectroscopy determined that the functionalization degree of the diethylamine-modified hyperbranched polyesters (LI, LII) increases from the second (56%) to third (81%) generation. The polynuclear complexes Cu(II)–LI and Cu(II)–LII were observed. A coordination site of the complexes contains paramagnetic fragment CuN2O2Solv2 where Solv is H2O or DMSO. The hyperbranched poly(ester amine) of the third generation have strong antifungal activity against Aspergillus fumigatus culture, and its metal complex is active against Candida albicans culture.

Bioconversion of oil palm frond to reducing sugar in solid state fermentation by applying water-fed strategy

Tones of Oil Palm Frond (OPF) waste from oil palm industries may affects the environment when released without proper treatment. Conversion of oil palm frond fiber into reducing sugar in the Solid State Fermentation (SSF) using Aspergillus fumigatus culture provide alternative for usage of these biomass. In this study, effect of the particle size of oil palm frond fiber and moisture content to the production of reducing sugar in SSF were investigated. Two particle size of OPF used are 100 and 250 μm while moiture level was set at 60, 70 and 80% (w/v). The fermentation process was carried out for 10 days. Effect of moisture was further analyzed by applying water-fed strategy. All fermentation using water-fed strategy yielded higher production of sugar than compared to others. Pmax of 4.446 g L-1 was achieved at 70% moisture level using 250 μm OPF while maximum Yp/s achieved was 0.442 (g g-1) in 80% moisture level using 250 μm OPF. Statistical analysis showed both particle size of oil palm frond fiber, 100 and 250 μm has no significant different towards production of reducing sugar in SSF by Aspergillus fumigatus. Statistically, moisture content of fiber does not significantly affect the production of reducing sugar but using water-fed strategy does gives significant difference. Sufficient moisture in SSF had increased the efficiency of fungus enzymatic system due to water holding capacity of substrate improved. Analysis two-way ANOVA suggest that presence of water fed has significant different towards production of reducing sugar in SSF and result in higher production of reducing sugar.

Improvement of fumigaclavine C production in a two-stage culture of Aspergillus fumigatus with molasses as a cost-effective ingredient

Fumigaclavine C (FC), which is produced by Aspergillus fumigatus, is a conidiation-associated ergot alkaloid with significant medical benefits. However, its application is restricted by low yields from submerged cultures. In this study, the technical feasibility of using molasses as a cost-effective ingredient for FC production in a two-stage culture of A. fumigatus was evaluated. The results indicated that molasses supplementation significantly enhanced FC accumulation by promoting conidiation and up-regulating hydroxymethylglutaryl-CoA reductase activity. Via the optimization of the two-stage process in the presence of molasses, FC production in shake flasks reached 226.9 mg/L, which was approximately three times that in the original medium (75.9 mg/L). The use of molasses as a cost-effective ingredient for FC fermentation was also successfully reproduced in a lab-scale bioreactor system in which the maximum FC production reached 215.0 mg/L. The FC production obtained in this study is the highest ever reported. This increased efficiency will enable large-scale production of FC and extend the application of molasses as a low-cost substrate for producing other conidiation-related secondary metabolites.

Secondary metabolites from Aspergillus fumigatus, an endophytic fungus from the liverwort Heteroscyphus tener (Steph.) Schiffn.

Three new metabolites, asperfumigatin (1), isochaetominine (10), and 8'-O-methylasterric acid (21), together with nineteen known compounds, were obtained from the culture of Aspergillus fumigatus, an endophytic fungus from the Chinese liverwort Heteroscyphus tener (Steph.) Schiffn. Their structures were established by extensive analysis of the spectroscopic data. The absolute configurations of 1 and 10 were determined by analysis of their respective CD spectra. Cytotoxicity of these isolates against four human cancer cell lines was also determined.

Study on a host–guest interaction of β-cyclodextrin with tebuconazole in water

In order to assess the effect of β-cyclodextrin (β-CD) on the tebuconazole (TEB) solubility in water, UV–VIS spectroscopic measurements of the tebuconazole solutions with β-cyclodextrin were examined, followed by a VP-ITC calorimetric titration of the solutions of tebuconazole with β-cyclodextrin. The stoichiometry of the tebuconazole–β-cyclodextrin inclusion complex formed and the physical and chemical parameters describing the complex were determined. The effect of fungicide included inside the macromolecule of β-cyclodextrin on the fungal culture (hyphae) of Aspergillus fumigatus was also assessed.

Bioconversion of tea polyphenols to bioactive theabrownins by Aspergillus fumigatus.

Theabrownins (TB) are water-soluble phenolic compounds associated with the various health benefits of Pu-erh tea, a post-fermented Chinese dark tea. This work reports on the production of theabrownins from infusions of sun-dried green tea leaves using a pure culture of Aspergillus fumigatus isolated from a solid-state Pu-erh tea fermentation. A theabrownins yield of 158gkg(-1) sun-dried green tea leaves was obtained in 6days at 45°C in an aerobic fermentation. In a 2l fermenter, the yield of theabrownins was 151gkg(-1) sun-dried green tea leaves in 48h of aerobic culture (45°C, 1 vvm aeration rate, 250rpm agitation speed). Extracellular polyphenol oxidase and peroxidase of A. fumigatus contributed to this bioconversion. Repeated batch fermentation process was used for producing theabrownins but was less productive than the batch process.

Extraction, partial purification and characterization of pectinases isolated from Aspergillus species cultured on mango ( Mangifera indica ) peels

Pectinase was produced from a culture of Aspergillus niger , Aspergillus fumigatus and Aspergillus flavus . Pectinase synthesis was achieved using mango ( Mangifera indica ) pectin extract as an inducer during pectinolytic fungi isolation while submerged fermentation process was carried out using ground mango peels as the sole carbon source. Substrate fermentation was evaluated within seven days by monitoring the pectinase activity every 24 h. The highest pectinase secretion was obtained from A. niger and A. fumigatus after 92 h (day 4) of incubation, while in A. flavus, it was after 120 h (day 5). Crude enzyme extracts from the three organisms were partially purified by a combination of ammonium sulphate precipitation and dialysis with an approximately two-fold purification of the pectinase and a yield of 5.4, 7.66 and 5.99% for A. niger, A. fumigatus and A. flavus , respectively after dialysis. The specific activities of 1.62, 1.79 and 1.86 U/mg for A. niger, A. fumigatus and A. flavus enzymes were calculated, respectively. Pectinase from A. niger and A. fumigatus had pH and temperature optima of 5.0 and 40°C, respectively, while that from A. flavus had pH and temperature optima of 5.0 and 45°C. The Michealis constant, K m and the maximum velocity, V max determined from Lineweaver-Burk plots of initial velocity data at different concentrations of the mango pectin extract were 0.357 mg/ml and 35.34 U; 0.156 mg/ml and 68.0 U; and 0.261 mg/ml and 60.61 U; for the enzymes from A. niger , A. fumigatus and A. flavus, respectively. The results suggest that mango peels can be used for value added synthesis of pectinase, an important enzyme with numerous biotechnological applications. Keywords: Mango peels, mango pectin extract, pectinase, Aspergillus niger , Aspergillus fumigatus, Aspergillus flavus , submerged fermentation, partial purification, characterization. African Journal of Biotechnology , Vol 13(24) 2445-2425

English

Pectinase was produced from a culture of Aspergillus niger, Aspergillus fumigatus and Aspergillus flavus. Pectinase synthesis was achieved using mango (Mangifera indica) pectin extract as an inducer during pectinolytic fungi isolation while submerged fermentation process was carried out using ground mango peels as the sole carbon source. Substrate fermentation was evaluated within seven days by monitoring the pectinase activity every 24 h. The highest pectinase secretion was obtained from A. niger and A. fumigatus after 92 h (day 4) of incubation, while in A. flavus, it was after 120 h (day 5). Crude enzyme extracts from the three organisms were partially purified by a combination of ammonium sulphate precipitation and dialysis with an approximately two-fold purification of the pectinase and a yield of 5.4, 7.66 and 5.99% for A. niger, A. fumigatus and A. flavus, respectively after dialysis. The specific activities of 1.62, 1.79 and 1.86 U/mg for A. niger, A. fumigatus and A. flavus enzymes were calculated, respectively. Pectinase from A. niger and A. fumigatus had pH and temperature optima of 5.0 and 40&deg;C, respectively, while that from A. flavus had pH and temperature optima of 5.0 and 45&deg;C. The Michealis constant, Km and the maximum velocity, Vmax determined from Lineweaver-Burk plots of initial velocity data at different concentrations of the mango pectin extract were 0.357 mg/ml and 35.34 U; 0.156 mg/ml and 68.0 U; and 0.261 mg/ml and 60.61 U; for the enzymes from A. niger, A. fumigatus and A. flavus, respectively. The results suggest that mango peels can be used for value added synthesis of pectinase, an important enzyme with numerous biotechnological applications. &nbsp; Key words: Mango peels, mango pectin extract, pectinase, Aspergillus niger, Aspergillus fumigatus, Aspergillus flavus, submerged fermentation, partial purification, characterization. &nbsp

Enhanced production of Fumigaclavine C in liquid culture of Aspergillus fumigatus under a two-stage process

Fumigaclavine C (FC) produced by Aspergillus fumigatus is a conidiation associated ergot alkaloid with strong anti-inflammatory activity. However, its wide application has been severely limited by low FC production from submerged culture. In this work, a novel two-stage culture process by combining shake culture with static culture was proposed to enhance the production of FC. After the process optimization, the FC production reached 62.7 mg/L, which was significantly higher than ever report. For scaling up this new culture process, the gas-liquid interfacial area per unit volume (Agas-liq) was identified as the key factor. The results showed that in a combined stirred-static bioreactor system, a maximum FC production (58.97 mg/L) was obtained at an Agas-liq value of 1.30 cm(2)/mL. These results demonstrated that two-stage culture is an efficient strategy to enhance FC production and the information obtained will be useful to production of this powerful bioactive compound on a large scale.