Mold Aspergillus Research Articles

Chronic Necrotizing Pulmonary Aspergillosis After SARS-CoV-2 Infection – A Case Report

Abstract Aspergillosis is a fungal infection, caused by the mould Aspergillus, most commonly Aspergillus fumigatus species. Chronic pulmonary aspergillosis after SARS Cov-2 infection is a rare presentation that is commonly misdiagnosed. The prolonged corticosteroid and antibiotic application and the pro-inflammatory state in COVID-19 patients predisposes to Aspergillus infection and its chronification. Surgery plays a pivotal role in cases with unclear diagnosis, ineffective medical therapy or when complications develop. We present a case of a 73-year-old woman with chronic pulmonary aspergillosis, developed after COVID-19 pneumonia. A right lateral muscle-sparing thoracotomy, right upper lobectomy and atypical resection of the 9th and 10th segments were performed.

Antifungal activity of the endophytic Aspergillus against Candida albicans

Medicinal plants remain a reliable source of bioactive compound principles known for their proven therapeutic abilities against various infectious diseases. Endophytes, microorganisms residing within plant tissues, hold promise for producing novel metabolites with potential medical applications. This study analyzes the antagonism of endophytic fungi Aspergillus sp., isolated from medicinal plants, and their extract against Candida albicans, and their effectiveness was compared with that of a medical treatment, Phanazol 1% ointment. After isolating, purifying, and identifying endophytic fungi from the medicinal plants Lavandula officinalis, Rosmarinus officinalis, Eucalyptus bicolor and Mentha piprita, a total of ten endophytic fungi were obtained. These included two yeasts (yeast and Rhodotorula sp.), as well as eight moulds (Aspergillus sp., Aspergillus niger, Nigrospora sp., Curvularia sp., Alternaria sp., Penicillium sp.), and sterile mycelium. All these fungi were tested for their antagonism against C. albicans; using the cross-streak and disk diffusion methods for yeasts and moulds respectively, with the measurement of the diameter of the growth inhibition zone of the culture. Only the strain Aspergillus sp. and its ethyl acetate extract exhibited good activity against C. albicans, with inhibition zone widths of 27.5 and 20.3 mm, respectively. Its effectiveness is comparable to that of Phanazol 1% ointment. The use of gas chromatography mass spectrometry (GC/MS) unveiled the metabolite profiles of Aspergillus sp., enabling the recognition of 10 bioactive compounds, with butanedioic acid, kojic acid, and Cyclo L-prolyl-L-valine being the major ones, constituting 45.1%, 23.1%, and 5.1% of the total, respectively. These compounds serve as valuable platform chemicals that can be transformed into various other useful chemicals with various applications in agriculture, pharmaceuticals, food, cosmetics, and the healthcare industry. In addition to refining the active substances within this extract, it has the potential to open doors for creating novel bio-sourced medications aimed at addressing resistant opportunistic fungal or bacterial infections.

Heterologous expression and characterization of salt-tolerant β-glucosidase from xerophilic Aspergillus chevalieri for hydrolysis of marine biomass.

A salt-tolerant exo-β-1,3-glucosidase (BGL_MK86) was cloned from the xerophilic mold Aspergillus chevalieri MK86 and heterologously expressed in A. oryzae. Phylogenetic analysis suggests that BGL_MK86 belongs to glycoside hydrolase family 5 (aryl-phospho-β-D-glucosidase, BglC), and exhibits D-glucose tolerance. Recombinant BGL_MK86 (rBGL_MK86) exhibited 100-fold higher expression than native BGL_MK86. rBGL_MK86 was active over a wide range of NaCl concentrations [0%-18% (w/v)] and showed increased substrate affinity for p-nitrophenyl-β-D-glucopyranoside (pNPBG) and turnover number (kcat) in the presence of NaCl. The enzyme was stable over a broad pH range (5.5-9.5). The optimum reaction pH and temperature for hydrolysis of pNPBG were 5.5 and 45°C, respectively. rBGL_MK86 acted on the β-1,3-linked glucose dimer laminaribiose, but not β-1,4-linked or β-1,6-linked glucose dimers (cellobiose or gentiobiose). It showed tenfold higher activity toward laminarin (a linear polymer of β-1,3 glucan) from Laminaria digitata than laminarin (β-1,3/β-1,6 glucan) from Eisenia bicyclis, likely due to its inability to act on β-1,6-linked glucose residues. The β-glucosidase retained hydrolytic activity toward crude laminarin preparations from marine biomass in moderately high salt concentrations. These properties indicate wide potential applications of this enzyme in saccharification of salt-bearing marine biomass.

A Multiplex PCR Melting-Curve-Analysis-Based Detection Method for the Discrimination of Five Aspergillus Species.

Aspergillus mold is a ubiquitously found, airborne pathogen that can cause a variety of diseases from mild to life-threatening in severity. Limitations in diagnostic methods combined with anti-fungal resistance render Aspergillus a global emerging pathogen. In industry, Aspergilli produce toxins, such as aflatoxins, which can cause food spoilage and pose public health risk issues. Here, we report a multiplex qPCR method for the detection and identification of the five most common pathogenic Aspergillus species, Aspergillus fumigatus, Aspergillus flavus, Aspergillus niger, Aspergillus terreus, and Aspergillus nidulans. Our approach exploits species-specific nucleotide polymorphisms within their ITS genomic regions. This novel assay combines multiplex single-color real time qPCR and melting curve analysis and provides a straight-forward, rapid, and cost-effective detection method that can identify five Aspergillus species simultaneously in a single reaction using only six unlabeled primers. Due to their unique fragment lengths, the resulting amplicons are directly linked to certain Aspergillus species like fingerprints, following either electrophoresis or melting curve analysis. Our method is characterized by high analytical sensitivity and specificity, so it may serve as a useful and inexpensive tool for Aspergillus diagnostic applications both in health care and the food industry.

Hematopoietic 12/15-lipoxygenase activity negatively contributes to fungal-associated allergic asthma.

Asthma is one of the most common non-communicable diseases in the world. Approximately 30% of severe cases are associated with fungal sensitization, often associated with allergy to the opportunistic mold Aspergillus fumigatus. Leukotrienes, immunopathogenic mediators derived from the metabolism of arachidonic acid (AA) by 5-lipoxygenase (5-LOX), are often elevated in severe asthma. As such, these mediators are FDA-approved therapeutic targets of the anti-asthmatic drugs Zileuton®/Zyflo® and Singulair®/Montelukast®. A second enzyme involved in AA metabolism is 12/15-lipoxygenase (12/15-LOX; Alox15). Here, C57BL/6 wild-type mice subjected to experimental fungal asthma had increased expression of Alox15 mRNA as well as increased levels of 12-HETE, a product of 12/15-LOX activity, in the lung when compared to naïve and vehicle-treated mice. Mice deficient in 12/15-LOX (Alox15-/-) demonstrated better lung function, as measured by airway hyperresponsiveness (AHR), during fungal asthma. Histological assessment revealed reduced inflammation in the lungs of Alox15-/- mice compared to WT mice, which was corroborated by flow cytometric analysis of multiple myeloid (eosinophils, neutrophils) and lymphoid (CD4+ T, γδ T) cell populations. This was further supported by decreased levels of specific chemokines that promote the recruitment of these cells. Likewise, type 1 and 2, but not type 17 cytokines, were significantly lower in the lungs of Alox15-/- mice. Bone marrow chimera studies revealed that the presence of 12/15-LOX in hematopoietic cells contributed to AHR during fungal asthma. Taken together, our data supports the hypothesis that hematopoietic-associated 12/15-LOX contributes to type 2 responses and exacerbation of allergic fungal asthma.

Preparation of Semi-Solid Dosage Forms Containing Psidium Guava Leaves Extract as Antimicrobial Preservative

Guava leaves extract has several medicinal activities as antimicrobial, antioxidant, anti-cancer and anti-inflammatory. Several studies have been proposed to use guava leaves extract in several preparations. However, applying guava leaves extract as a preservative in a pharmaceutical product has not been studied previously. In the present work, Guava leaves extract was used as natural preservative instead of chemical ones (methyl and propyl paraben). Guava leaves were extracted with ethanol 95% as extraction solvent with a percentage yield of 20%. This extract was tested by HPLC; several phenolic compounds were detected. To determine the ability to use this extract as a preservative, antimicrobial effectiveness test (preservative efficacy test) was conducted for extracted powder in purified water at different concentrations (0.5, 1.0, 1.5, and 2.0 % w/w), against three bacteria, one gram-positive (S. aureus), two gram-negative (P. aeruginosa, E. coli) and two fungi: one yeast (Candida albicans) and one mold (Aspergillus (Niger) brasiliensis). Results showed that 2% was effective and passed the test of preservative effectiveness test; this concentration was used in semi-solid pharmaceutical products. For this purpose, six different preparations (Ketoconazole shampoo, Clotrimazole Cream, Permethrin Cream, Gentamycin Cream, Ibuprofen gel, Indomethacin emulgel) were used. Chemical preservative was replaced by natural one (guava leaves extract powder) to serve as a preservative. Three out of the six preparations (Ketoconazole shampoo, Clotrimazole Cream, Ibuprofen gel,) passed preservative efficacy test. These preparations were fully checked-up by chemical (drug content,) physical (odor, physical appearance) and biological test (total count test) for three months at accelerated conditions. These results confirmed that these pharmaceutical preparations were stable and effective. As a conclusion Guava leaves extract can be used at 2% as natural preservative instead of chemical ones which have adverse side effects on human health.

Possibilities of CT in the diagnosis of pulmonary aspergillosis in patients with COVID-19 (literature review)

Aim: to analyze the latest research in the field of CT diagnostics of pulmonary aspergillosis in patients with COVID-19 according to domestic and foreign literature.Results. Pulmonary aspergillosis is one of the most common life-threatening fungal diseases that is caused by inhalation of Aspergillus mold spores and affects all parts of the respiratory system. In recent years, there has been a sharp increase in cases of invasive mycotic processes. During the COVID-19 pandemic, according to a number of authors, the incidence of pulmonary aspergillosis reached 34.4%, and the mortality rate was 74%. The World Health Organization calls for “minimizing the disproportionate use of corticosteroids and antibiotics for mild and moderate cases”. She explained that corticosteroids in severe cases do save lives, but their massive use can cause fungal infections. This complication is increasingly common in patients after corticosteroid and antibiotic therapy, with immunodeficiency states, as well as those treated in intensive care units. The COVID-19 pandemic has created additional challenges in the diagnosis, treatment and prevention of fungal complications. The clinical and radiographic features of fungal infections are largely similar to typical pulmonary manifestations of COVID-19. A comparative analysis of CT signs of various forms of pulmonary aspergillosis before and during the COVID-19 pandemic was carried out according to the literature. The main CT symptoms of mycotic lung lesions among patients who have undergone COVID-19 have been identified. Currently, computed tomography is crucial, first of all, to detect and monitor complications of COVID-19, including the addition of a secondary fungal infection.Conclusion. To make a conclusion about mycosis in the early stages, special vigilance and high qualification of the radiologist are required, which in turn will contribute to the timely appointment of antifungal therapy, which will reduce morbidity and mortality.

Determination of aspergillosis of the female genital organs in women with immunodeficiency

Background. Morphological diagnosis of aspergillosis sometimes poses great difficulties due to the fact that molds of the genus Aspergillus are very sensitive to the use of antimicrobial therapy and change under its influence. In this regard, we provide information about aspergillosis of the female genital organs, discovered accidentally during an autopsy. The purpose of the study is to conduct a complex pathomorphological analysis of biopsy, operative and sectional gynecological material of women with secondary immunodeficiency conditions, for early diagnosis of aspergillosis (which is the key to a favorable outcome of the disease), differential diagnosis and detection of invasive forms of aspergillosis. Materials & Methods. We analyzed the autopsies of 4,796 women who died in Odesa region from 2009 to 2022. Autopsies revealed 7 cases of aspergillosis with damage to the female reproductive system. All of them were found in women who had confirmed HIV infection. In 5 cases, women additionally suffered from chronic alcoholism. Results. Based on the results of these cases, it can be stated that the detection of characteristic mycelium in the preparations is a reliable sign of aspergillosis, but their absence does not deny this disease. Aspergillus hyphae change as the process develops: they can be weakly stained, and with dystrophic phenomena and the presence of necrotic fragments of mycelium, it can be almost colorless when using hematoxylin and eosin. While necrosis, tissue detritus is observed in the center of the granuloma, fibrous structures, fungal hyphae are formed perifocally. To confirm the diagnosis, it is important to detect granulomatous productive inflammation, conidial heads, inflammatory infiltration, microabscesses, foci of calcifications. However, against the background of immunodeficiency, it is not always possible to see the "classic morphological picture" of aspergillosis. In addition, detection of characteristic mycelium is possible only in a proportion of patients. Conclusions. When examining the autopsy material of deceased women with immunodeficiency states, attention should be paid to the specific etiology of inflammatory processes of the reproductive system and the nature of the inflammatory process, the presence of Aspergillus hyphae and mycelia. The presence of morphological changes characteristic of aspergillosis allows to conduct a diagnostic search in the right direction. Detection of an invasive form of aspergillosis may indicate the presence of an immunodeficient state. Keywords: aspergillosis, female reproductive system, immunodeficiency state, HIV infection, uterus, mammary gland, drug addiction.

Identification of triazenyl indoles as inhibitors of fungal fatty acid biosynthesis with broad-spectrum activity

Rising drug resistance among pathogenic fungi, paired with a limited antifungal arsenal, poses an increasing threat to human health. To identify antifungal compounds, we screened the RIKEN natural product depository against representative isolates of four major human fungal pathogens. This screen identified NPD6433, a triazenyl indole with broad-spectrum activity against all screening strains, as well as the filamentous mold Aspergillus fumigatus. Mechanistic studies indicated that NPD6433 targets the enoyl reductase domain of fatty acid synthase 1 (Fas1), covalently inhibiting its flavin mononucleotide-dependent NADPH-oxidation activity and arresting essential fatty acid biosynthesis. Robust Fas1 inhibition kills Candida albicans, while sublethal inhibition impairs diverse virulence traits. At well-tolerated exposures, NPD6433 extended the lifespan of nematodes infected with azole-resistant C.albicans. Overall, identification of NPD6433 provides a tool with which to explore lipid homeostasis as a therapeutic target in pathogenic fungi and reveals a mechanism by which Fas1 function can be inhibited.

Investigation of the Antimicrobial and Physico-Mechanical Properties of Nature-Friendly Nanosilver-Loaded Pig Lining Leather Prepared Using Exhaustion Method

The natural hydrophilic property of leather makes it a possible growing substrate for microorganisms. Microorganisms such as bacteria, yeasts, and molds frequently grow on shoe lining leather, causing skin diseases, unpleasant odors, and discomfort for the wearers. To address these issues, herein, bio-synthesized silver nanoparticles from Piper betle L. leaf extract were applied as an antimicrobial agent to pig leather via the exhaustion method. The characterization of nanosilver-treated pig leather was performed using colorimetry, scanning electron microscopy (SEM), energy-dispersive X-ray spectroscopy (EDX), atomic absorption spectroscopy (AAS), and FTIR techniques. Evaluation of antimicrobial efficacy of the nanosilver-treated leather was both qualitatively and quantitatively assessed against two bacteria (Escherichia coli and Staphylococcus aureus) and two fungi (a yeast Candida albicans and a mold Aspergillus niger) in accordance with AATCC TM90, AATCC TM30, and ISO 16187:2013 standards. The results indicated that the nanosilver was immobilized on the surface of collagen fibers as well as within the collagen matrix of the pig leather. The treated leather exhibited highly effective antibacterial and antifungal activities against all tested microorganisms, and the inhibition increased with an increase in the initial nanosilver concentration in the treated solution. Furthermore, the exhaustion technique used for the antimicrobial treatment of pig leather had no negative effects on its physico-mechanical properties, and it met the standard requirements of ISO 20882:2007 for shoe upper lining. Therefore, based on the efficient antimicrobial and suitable physico-mechanical properties, nanosilver-treated pig leather adapts the criteria for making hygienic shoe upper lining.

Antibacterial and Antifungal Fabrication of Natural Lining Leather Using Bio-Synthesized Silver Nanoparticles from Piper Betle L. Leaf Extract.

Leather is often used to make comfortable shoes due to its soft and breathable nature. However, its innate ability to retain moisture, oxygen and nutrients renders it a suitable medium for the adsorption, growth, and survival of potentially pathogenic microorganisms. Consequently, the intimate contact between the foot skin and the leather lining surface in shoes, which are subject to prolonged periods of sweating, may result in the transmission of pathogenic microorganisms and cause discomfort for the wearer. To address such issues, we modified pig leather with silver nanoparticles (AgPBL) that were bio-synthesized from Piper betle L. leaf extract as an antimicrobial agent via the padding method. The evidence of AgPBL embedded into the leather matrix, leather surface morphology and element profile of AgPBL-modified leather samples (pLeAg) was investigated using colorimetry, SEM, EDX, AAS and FTIR analyses. The colorimetric data confirmed that the pLeAg samples changed to a more brown color with higher wet pickup and AgPBL concentration, owing to the higher quantity of AgPBL uptake onto the leather surfaces. The antibacterial and antifungal activities of the pLeAg samples were both qualitatively and quantitatively evaluated using AATCC TM90, AATCC TM30 and ISO 16187:2013 test methods, approving a good synergistic antimicrobial efficiency of the modified leather against Escherichia coli and Staphylococcus aureus bacteria, a yeast Candida albicans and a mold Aspergillus niger. Additionally, the antimicrobial treatments of pig leather did not negatively impact its physico-mechanical properties, including tear strength, abrasion resistance, flex resistance, water vapour permeability and absorption, water absorption and desorption. These findings affirmed that the AgPBL-modified leather met all the requirements of upper lining according to the standard ISO 20882:2007 for making hygienic shoes.

The Diagnostic Value of Delayed-Type Reactions to Perennial Aeroallergens for Atopic Disease.

Background: Delayed-type reactions to aeroallergens have been observed, however, their clinical significance continues to be debated. Objective: We assessed the prevalence and significance of delayed-type reactions to aeroallergens in atopic patients. Methods: Retrospective study including 266 patients with history or evidence of atopic disease (atopic dermatitis [AD], allergic rhinitis, and/or allergic asthma) and tested via either the intradermal skin test (IDT) or atopy patch test for common aeroallergens, specifically house dust mites (Dermatophagoides farinae, Dermatophagoides pteronyssinus) and perennial molds (Aspergillus fumigatus, Penicillium notatum). All patients were tested via IDT with both immediate (15 minute) and delayed (2 and 4 days) readings. Delayed reading was considered positive if the IDT injection site demonstrated at least 5 mm induration 48 hours after inoculation. Results: In total, 195 (73.3%) patients demonstrated an immediate-type reaction, whereas 118 (44.4%) had a delayed-type reaction. In total, 75 (28.2%) patients experienced both immediate- and delayed-type reactions, 43 (16.2%) reacted delayed-type only, and 85.3% of delayed-type reactions to individual aeroallergens were associated with eczematous lesions predominantly in air-exposed areas. Conclusion: Delayed-type reactions to aeroallergens are prevalent and clinically significant as a component of extrinsic AD and atopic diseases. The data support delayed reading of the IDT to guide diagnosis and management in these patients.

The effect of Aspergillus luchuensis pectin methylesterase genes pmeA and pmeB on methanol production in sweet potato shochu.

To reduce the methanol content in sweet potato shochu, we studied the pectin methylesterase genes of the shochu-koji mold Aspergillus luchuensis. We found the following three homologs of pectin methyleseterase in the genome of A. luchuensis: pmeA, pmeB, and pmeC. Using pectin as a substrate, the methanol-producing activity of the recombinant of each gene expressed in A. luchuensis was examined and found to be present in recombinant PmeA and PmeB. Additionally, small-scale fermentation of sweet potato shochu using disruptions of pmeA and pmeA-pmeB in A. luchuensis (∆pmeA and ∆pmeApmeB) resulted in significant reduction of the methanol content. Taken together, we revealed that the A. luchuensis pmeA gene was mainly involved in methanol production in sweet potato shochu.

An efficient Agrobacterium-mediated system based on the pyrG auxotrophic marker for recombinant expression in the filamentous fungus Penicillium rubens

This work aimed to construct a versatile, effective, and food-grade Agrobacterium tumefaciens-mediated transformation (ATMT) system for recombinant expression in the filamentous fungus Penicillium rubens (also known as Pencillium chrysogenum). In this study, the wild-type P. chrysogenum VTCC 31172 strain was re-classified as P. rubens by a multilocus sequencing analysis. Further, the pyrG gene required for uridine/uracil biosynthesis was successfully deleted in the VTCC 31172 strain by homologous recombination to generate a stable uridine/uracil auxotrophic mutant (ΔpyrG). The growth of the P. rubens ΔpyrG strain could be restored by uridine/uracil supplementation, and a new ATMT system based on the uridine/uracil auxotrophic mechanism was established for this strain. The optimal ATMT efficiency could reach 1750 transformants for 106 spores (equivalent to 0.18%). In addition, supplementation of uridine/uracil at the concentrations of 0.005-0.02% during the co-cultivation process significantly promoted transformation efficiency. Especially, we demonstrated that the pyrG marker and the amyB promoter from the koji mold Aspergillus oryzae were fully functional in P. rubens ΔpyrG. Expression of the DsRed reporter gene under the regulation of the A. oryzae amyB promoter lighted up the mycelium of P. rubens with a robust red signal under fluorescence microscopy. Furthermore, genomic integration of multiple copies of the Aspergillus fumigatus phyA gene under the control of the amyB promoter significantly enhanced phytase activity in P. rubens. The ATMT system developed in our work provides a safe genetic platform for producing recombinant products in P. rubens without using drug resistance markers.

Rapid but narrow - Evolutionary adaptation and transcriptional response of Drosophila melanogaster to toxic mould.

Insects have adapted to a multitude of environmental conditions, including the presence of xenobiotic noxious substances. Environmental microorganisms, particularly rich on ephemeral resources, employ these noxious chemicals in a chemical warfare against predators and competitors, driving co-evolutionary adaptations. In order to analyse how environmental microbes may be driving such evolutionary adaptations, we experimentally evolved Drosophila melanogaster populations by exposing larvae to the toxin-producing mould Aspergillus nidulans that infests the flies' breeding substrate. To disentangle the effects of the mycotoxin Sterigmatocystin from other substrate modifications inflicted by the mould, we used the following four selection regimes: (i) control without fungus, (ii) A. nidulans wild type, (iii) a mutant of A. nidulans ΔlaeA with impaired toxin production, (iv) synthetic Sterigmatocystin. Experimental evolution was carried out in five independent D. melanogaster populations each, for a total of 11 generations. We further combined our evolution experiment with transcriptome analysis to identify evolutionary shifts in gene expression due to the selection regimes and mould confrontation. Populations that evolved in presence of the toxin-producing mould or the pure mycotoxin rapidly adapted to the respective conditions and showed higher viability in subsequent confrontations. Yet, mycotoxin-selected populations had no advantage in A. nidulans wild type confrontation. Moreover, distinctive changes in gene expression related to the selection-regime contrast were only associated with the toxin-producing-fungus regime and comprised a narrow set of genes. Thus, it needs the specific conditions of the selection agent to enable adaptation to the fungus.

Functionalization of silk with chitosan and Rubia cordifolia L. dye extract for enhanced antimicrobial and ultraviolet protective properties

Natural dyes have attained the interest of scientists over the last decade due to their functional activities in addition to their primary role of color impartation on textile materials. This study is the first report on the functionalization of silk with antimicrobial and ultraviolet protective properties using a dye extract from the roots of Rubia cordifolia L. in conjunction with chitosan as a bio-mordant. Alum mordant was also used in the dyeing process alongside chitosan for comparison purposes. Fourier transform infrared spectroscopy and ultraviolet-visible spectroscopy were utilized to determine the anthraquinone-based components of the dye extract. Furthermore, the structure and surface morphology of silk fibers before and after dyeing were investigated. The dyed silk samples exhibited good fastness against washing, rubbing, and lighting. The chitosan mordanted-dyed silk presented remarkable antimicrobial properties with maximum inhibitions of 96.71% and 99.08% against Escherichia coli and Staphylococcus aureus, respectively, and only 2.3% spore germination of white mold ( Aspergillus spp.). Likewise, an excellent ultraviolet protection factor was achieved in the chitosan modified-dyed silk. The findings of this study demonstrated the potential of the Rubia cordifolia L. extract as a functional dye, as well as the advanced properties of chitosan in increasing the fastness and functional performance for the dyed silk compared to alum mordant.

Random mutagenesis of koji mold (Aspergillus oryzae) to enhance the catalytic efficiency and thermostability of glucoamylase

Random mutagenesis of koji mold (Aspergillus oryzae) to enhance the catalytic efficiency and thermostability of glucoamylase

COVID-19-Associated Pulmonary Aspergillosis Isolates Are Genomically Diverse but Similar to Each Other in Their Responses to Infection-Relevant Stresses.

Secondary infections caused by the pulmonary fungal pathogen Aspergillus fumigatus are a significant cause of mortality in patients with severe coronavirus disease 19 (COVID-19). Even though epithelial cell damage and aberrant cytokine responses have been linked to susceptibility to COVID-19-associated pulmonary aspergillosis (CAPA), little is known about the mechanisms underpinning copathogenicity. Here, we analyzed the genomes of 11 A. fumigatus isolates from patients with CAPA in three centers from different European countries. CAPA isolates did not cluster based on geographic origin in a genome-scale phylogeny of representative A. fumigatus isolates. Phenotypically, CAPA isolates were more similar to the A. fumigatus A1160 reference strain than to the Af293 strain when grown in infection-relevant stresses, except for interactions with human immune cells wherein macrophage responses were similar to those induced by the Af293 reference strain. Collectively, our data indicate that CAPA isolates are genomically diverse but are more similar to each other in their responses to infection-relevant stresses. A larger number of isolates from CAPA patients should be studied to better understand the molecular epidemiology of CAPA and to identify genetic drivers of copathogenicity and antifungal resistance in patients with COVID-19. IMPORTANCE Coronavirus disease 2019 (COVID-19)-associated pulmonary aspergillosis (CAPA) has been globally reported as a life-threatening complication in some patients with severe COVID-19. Most of these infections are caused by the environmental mold Aspergillus fumigatus, which ranks third in the fungal pathogen priority list of the WHO. However, little is known about the molecular epidemiology of Aspergillus fumigatus CAPA strains. Here, we analyzed the genomes of 11 A. fumigatus isolates from patients with CAPA in three centers from different European countries, and carried out phenotypic analyses with a view to understanding the pathophysiology of the disease. Our data indicate that A. fumigatus CAPA isolates are genomically diverse but are more similar to each other in their responses to infection-relevant stresses.

Simultaneous detection of Aspergillus moulds and aflatoxin B1 contamination in rice by laser induced fluorescence spectroscopy

Mould and aflatoxin contamination remains a serious threat to food quality and safety. In this study, simultaneous detection of Aspergillus moulds and aflatoxin B1 (AFB1) contamination in rice was investigated by laser induced fluorescence (LIF) technology for the first time. Different Aspergillus strains belong to toxigenic and non-toxigenic species were artificially inoculated in rice samples and storage for a period of time to evolve into various mould and AFB1 infection levels. Spectral analysis and principal component analysis (PCA) indicated that mould activity could be monitored by LIF spectroscopy. Subsequently, three classification methods based on linear and non-linear algorithms were established to classify rice samples in terms of the type of infected mould, mould infection level as well as AFB1 concentration. The best correct classified rate obtained achieved 97% with false positive rates less than 2%. In addition, the discrimination towards rice flour samples performed better than kernel samples. These findings verify the potential of LIF spectroscopy as a rapid and non-destructive way for sensitive and effective detection of mould and aflatoxin contamination in grains without any complicated pre-processing.

Aspergillus-Specific IgG Antibodies are Associated With Fungal-Related Complications and Chronic Lung Allograft Dysfunction After Lung Transplantation.

Fungal exposure and sensitization negatively affect outcomes in various respiratory diseases, however, the effect of fungal sensitization in lung transplant (LTx) recipients is still unknown. We performed a retrospective cohort study of prospectively collected data on circulating fungal specific IgG/IgE antibodies, and their correlation with fungal isolation, chronic lung allograft dysfunction (CLAD) and overall survival after LTx. 311 patients transplanted between 2014 and 2019 were included. Patients with elevated Aspergillus fumigatus or Aspergillus flavus IgG (10%) had more mold and Aspergillus species isolation (p = 0.0068 and p = 0.0047). Aspergillus fumigatus IgG was specifically associated with Aspergillus fumigatus isolation in the previous or consecutive year (AUC 0.60, p = 0.004 and AUC 0.63, p = 0.022, respectively). Elevated Aspergillus fumigatus or Aspergillus flavus IgG was associated with CLAD (p = 0.0355), but not with death. Aspergillus fumigatus, Aspergillus flavus or Aspergillus niger IgE was elevated in 19.3% of patients, but not associated with fungal isolation, CLAD or death. Mold isolation and Aspergillus species isolation from respiratory cultures were associated with CLAD occurrence (p = 0.0011 and p = 0.0005, respectively), and Aspergillus species isolation was also associated with impaired survival (p = 0.0424). Fungus-specific IgG could be useful in long-term follow-up post-LTx, as a non-invasive marker for fungal exposure, and thus a diagnostic tool for identifying patients at risk for fungal-related complications and CLAD.