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Second generation biorefineries play an important role in the production of renewable energy and fuels, utilizing forest and agro-industrial residues and by-products as raw materials. The integration of novel bioproducts, such as: xylitol, ß-carotene, xylooligosaccharides, and biopigments into the biorefinery's portfolio can offer economic benefits in the valorization of lignocellulosic materials, particularly cellulosic and hemicellulosic fractions. Fungal biopigments, known for their additional antioxidant and antimicrobial properties, are appealing to consumers and can have applications in various industrial sectors, including food and pharmaceuticals. The use of lignocellulosic materials as carbon and nutrient sources for the growth medium helps to reduce production costs, increasing the competitiveness of fungal biopigments in the market. In addition, the implementation of biopigment production in biorefineries allows the utilization of underutilized fractions, such as hemicellulose, for value-added bioproducts. This study deals with the potential of fungal biopigments production in second generation biorefineries in order to diversify the produced biomolecules together with energy generation. A comprehensive and critical review of the recent literature on this topic has been conducted, covering the major possible raw materials, general aspects of second generation biorefineries, the fungal biopigments and their potential for incorporation into biorefineries.

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In this comprehensive study, we delved into the capabilities of five fungal strains: Aspergillus flavus, Aspergillus niger, Penicillium chrysogenum, Penicillium glabrum, and Penicillium rubens (the latter isolated from heavy crude oil [HCO]) in metabolizing HCO as a carbon source. Employing a meticulously designed experimental approach, conducted at room temperature (25 °C), we systematically explored various culture media and incubation periods. The results unveiled the exceptional resilience of all these fungi to HCO, with A. flavus standing out as the top performer. Notably, A. flavus exhibited robust growth, achieving a remarkable 59.1% expansion across the medium's surface, accompanied by distinctive macroscopic traits, including a cottony appearance and vibrant coloration. In an effort to further scrutinize its biotransformation prowess, we conducted experiments in a liquid medium, quantifying CO2 production through gas chromatography, which reached its zenith at day 30, signifying substantial bioconversion with a 38% increase in CO2 production. Additionally, we monitored changes in surface tension using the Du Noüy ring method, revealing a reduction in aqueous phase tension from 72.3 to 47 mN/m. This compelling evidence confirms that A. flavus adeptly metabolizes HCO to fuel its growth, while concurrently generating valuable biosurfactants. These findings underscore the immense biotechnological potential of A. flavus in addressing challenges related to HCO, thereby offering promising prospects for bioremediation and crude oil bioupgrading endeavors.

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SUMMARYThe endoplasmic reticulum (ER) is one of the most extensive organelles in eukaryotic cells. It performs crucial roles in protein and lipid synthesis and Ca2+ homeostasis. Most information on ER types, functions, organization, and domains comes from studies in uninucleate animal, plant, and yeast cells. In contrast, there is limited information on the multinucleate cells of filamentous fungi, i.e., hyphae. We provide an analytical review of existing literature to categorize different types of ER described in filamentous fungi while emphasizing the research techniques and markers used. Additionally, we identify the knowledge gaps that need to be resolved better to understand the structure-function correlation of ER in filamentous fungi. Finally, advanced technologies that can provide breakthroughs in understanding the ER in filamentous fungi are discussed.

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IMPORTANCE: Sexual reproduction allows eukaryotic organisms to produce genetically diverse progeny. This process relies on meiosis, a reductional division that enables ploidy maintenance and genetic recombination. Meiotic differentiation also involves the renewal of cell functioning to promote offspring rejuvenation. Research in the model fungus Podospora anserina has shown that this process involves a complex regulation of the function and dynamics of different organelles, including peroxisomes. These organelles are critical for meiosis induction and play further significant roles in meiotic development. Here we show that PEX13-a key constituent of the protein conduit through which the proteins defining peroxisome function reach into the organelle-is subject to a developmental regulation that almost certainly involves its selective ubiquitination-dependent removal and that modulates its abundance throughout meiotic development and at different sexual differentiation processes. Our results show that meiotic development involves a complex developmental regulation of the peroxisome protein translocation system.

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Background: Onychomycoses are fungal infections that can be seen in any component of the nail unit, including the matrix, bed, and plate, and are caused by dermatophyte fungi, non-dermatophyte fungi, and yeasts. This disease affects approximately 1 to 8% of the general population and occurs in approximately 19 to 51.9% of the patients on hemodialysis. The high incidence of onychomycosis in patients on hemodialysis is associated, mainly, with immunologic deficits and histological changes caused by uremia. Methods: Adult patients of the São Francisco University Hospital Hemodialysis Center were included. The following characteristics were evaluated: age, sex, body mass index, comorbidity, and household location. All patients were clinically evaluated and those with suspected onychomycosis had subungual debris of the affected nail plate collected for the direct mycological examination and fungal culture. The onychomycosis severity for those patients with a positive result in the fungal culture examination was evaluated using the Onychomycosis Severity Index system. Results: The study included 151 patients, and 70 out of the 151 patients (46.4%) showed nail alteration, and among them, 31 out of the 70 patients (44.3%) had the onychomycosis diagnosis confirmed by direct mycological examination. The pathogens observed in the patients were Trichophyton rubrum [8 out of 31 (25.8%)], Trichophyton mentagrophytes [7 out of 31 (22.6%)], Scytalidium spp. [6 out of 31 (19.4%)], Candida spp. [2 out of 31 (6.45%)], Rhodotorula spp. [1 out of 31 (3.2%)], Candida albicans [1/31 (3.2%)], Penicillium marneffei [1 out of 31 (3.22%)], and T. rubrum and Rhodotorula spp. [1 out of 31 (3.2%)]. Three participants presented negative results in the culture examination, and one did not allow the collection of material for the examination. The nail involvement severity score for the majority of them was severe [23 out of 27 (85.2%)], and only 1 out of the 27 (3.7%) and 3 out of the 27 (1.1%) patients presented moderate and mild scores, respectively. The distal subungual onychomycosis occurred in 12 out of 27 (44.4%) patients, a mixed pattern occurred in 14 out of 27 (51.9%) patients, and, white superficial occurred in only 1 out of 27 (3.7%) patients. In the bivariate analysis, a higher risk of onychomycosis was associated with the male sex [23/31 (74.2%) vs. 56/120 (46.7%); OR = 3.286 (95%CI = 1.362 to 7.928)] and obesity [8/31 (25.8%) vs. 12/120 (10.0%); OR = 3.130 (95%CI = 1.150 to 8.521)]. Patients with diabetes mellitus were more susceptible to onychomycosis attacks (p-value = 0.049; 16 out of 31 (51.6%) vs. 40 out of 120 (33.3%); however, OR was 2.133 (95%CI = 0.959 to 4.648). The patients with onychomycosis were older but without a significant difference between the groups (p-value = 0.073; 66 years old vs. 58 years old). The multivariable model using the logistic regression (backward model) confirmed our results and was able to predict (81.5%) the onychomycosis-positive diagnosis (chi-square = 27.73; p-value <0.001). The age [OR = 1.036; 95%CI = 1.004 to 1.069], male sex [OR = 5.746; 95%CI = 2.057 to 16.046], and presence of obesity [OR = 4.800; 95%CI = 1.435 to 16.055] were positive and significant in predicting the onychomycosis-positive diagnosis. Conclusion: In our study, onychomycosis in patients on hemodialysis was associated with a great variety of microorganisms, mainly Trichophyton species. The nail involvement severity score for the majority of patients was severe, and distal subungual onychomycosis and mixed pattern onychomycosis were the most prevalent clinical types. The main risk factors associated with onychomycosis were male sex, older age, and the presence of obesity.

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The present study aimed to investigate the antifungal activity of citronellal (CIT) against clinical isolates of T. rubrum and to show the possible mechanism of action involved. The antifungal potential of CIT was evaluated from the Minimum Inhibitory Concentration (MIC), Minimum Fungicide Concentration (MFC) and assays with ergosterol and sorbitol, to elucidate the possible mechanisms of action, and molecular docking. MIC and MFC values ranged from 4 to 512 µg/mL. Regarding the mechanism of action, the monoterpene demonstrated interaction with fungal ergosterol. In addition, it is possible to observe that CIT acts on crucial enzymes for the biosynthesis and maintenance of the fungal cell membrane, due to the ability of the monoterpene to bind to CYP51. The results obtained in this research demonstrate that CIT has the potential to become, in the future, a product for the treatment of dermatophytosis.

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The use of phosphate-solubilizing fungi in coffee cultivation is an alternative to the use of traditional fertilizers. The objective of this study was to analyze the mechanisms involved in the phosphorus solubilization of fungal strains and to evaluate the effect of a phosphate-solubilizing strain on coffee plants. For this, phosphorus-solubilizing fungal strains were selected for evaluation of their solubilization potential and phosphatase activity. Coffee plants were inoculated in the field with a phosphate-solubilizing strain, and the soil and foliar soluble phosphorus contents, as well as coffee bean yield, were quantified. Of the 151 strains analyzed, Sagenomella diversispora, Penicillium waksmanii, and Penicillium brevicompactum showed the highest solubilization. Aspergillus niger and P. waksmanii presented the highest soluble phosphorus values; however, P. brevicompactum showed the highest phosphatase activity. The P. brevicompactum strain inoculated on the coffee plants did not favor the foliar phosphorus content but increased the soil soluble phosphorus content in two of the coffee plantations. The plants inoculated with the phosphate-solubilizing strain showed an increase in coffee bean weight on all plantations, although this increase was only significant in two of the three selected coffee plantations.

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Interdisciplinary studies on cultural heritage artworks provide efficient solutions to control fungal growth and the negative effects of biodeterioration. In this study, we aimed to identify the population of filamentous fungi colonizing an engraving by the Dutch painter Rembrandt, whose conservation status was compromised and showed visible stains of biodeterioration. Microbiological techniques, such as cultivation-dependent approaches and molecular biology, have been used to identify fungal populations. In addition, the anaerobic atmosphere technique and eco-friendly antifungal agents, such as essential oils (EOs) of Curcuma longa, Thymus vulgaris, and Melaleuca alternifolia, were tested against the metabolically active fungal population Cladoposporium spinulosum. Furthermore, in vitro assays revealed that the interaction between the fungal strains and EO was positive, inhibiting the growth of these fungi, and the EOs from T. vulgaris and M. alternifolia showed low minimum inhibitory concentration values. Exposure to anaerobic conditions for 35 days was effective in the total elimination of isolated fungal strains. In conclusion, this study demonstrated the effectiveness of a nondestructive technique for artwork on engraving colonized by fungal strains and using EO as an alternative to toxic antifungals used in conventional treatments in artworks. Thus, this interdisciplinary study involving applied microbiology and botanical and preventive conservation presents a tool to control microbial growth while maintaining artwork integrity.

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Filamentous fungi develop intricate hyphal networks that support mycelial foraging and transport of resources. These networks have been analyzed recently using graph theory, enabling the development of models that seek to predict functional traits. However, attention has focused mainly on mature colonies. Here, we report the extraction and analysis of the graph corresponding to Trichoderma atroviride mycelia only a few hours after conidia germination. To extract the graph for a given mycelium, a mosaic conformed of multiple bright-field, optical microscopy images is digitally processed using freely available software. The resulting graphs are characterized in terms of number of nodes and edges, average edge length, total mycelium length, hyphal growth unit, maximum edge length and mycelium diameter, for colonies between 8 h and 14 h after conidium germination. Our results show that the emerging hyphal network grows first by hyphal elongation and branching, and then it transitions to a stage where hyphal-hyphal interactions become significant. As a tangled hyphal network develops with decreasing hyphal mean length, the mycelium maintains long (∼2 mm) hyphae-a behavior that suggests a combination of aggregated and dispersed architectures to support foraging. Lastly, analysis of early network development in Podospora anserina reveals striking similarity with T. atroviride, suggesting common mechanisms during initial colony formation in filamentous fungi.

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Lignocellulosic biomass is a promising alternative for producing biofuels, despite its recalcitrant nature. There are microorganisms in nature capable of efficiently degrade biomass, such as the filamentous fungi. Among them, Aspergillus fumigatus var. niveus (AFUMN) has a wide variety of carbohydrate-active enzymes (CAZymes), especially hydrolases, but a low number of oxidative enzymes in its genome. To confirm the enzymatic profile of this fungus, this study analyzed the secretome of AFUMN cultured in sugarcane bagasse as the sole carbon source. As expected, the secretome showed a predominance of hydrolytic enzymes compared to oxidative activity. However, it is known that hydrolytic enzymes act in synergy with oxidative proteins to efficiently degrade cellulose polymer, such as the Lytic Polysaccharide Monooxygenases (LPMOs). Thus, three LPMOs from the fungus Thermothelomyces thermophilus (TtLPMO9D, TtLPMO9H, and TtLPMO9O) were selected, heterologous expressed in Aspergillus nidulans, purified, and used to supplement the AFUMN secretome to evaluate their effect on the saccharification of sugarcane bagasse. The saccharification assay was carried out using different concentrations of AFUMN secretome supplemented with recombinant T. thermophilus LPMOs, as well as ascorbic acid as reducing agent for oxidative enzymes. Through a statistic design created by Design-Expert software, we were able to analyze a possible cooperative effect between these components. The results indicated that, in general, the addition of TtLPMO9D and ascorbic acid did not favor the conversion process in this study, while TtLPMO9O had a highly significant cooperative effect in bagasse saccharification compared to the control using only AFUMN secretome.

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