Filamentous fungi have potential as an alternative protein source for use in aquafeeds due to their high protein content and bioactive components. Paecilomyces variotii (PEKILO® mycoprotein) is a ...β-glucan- and nucleotide-rich filamentous fungi with a crude protein (CP) content of 60–70%. A 9-week feeding trial was conducted to evaluate the effects of P. variotii in diets for Atlantic salmon (initial average body weight of 24 g fish−1) reared in freshwater. Four experimental diets were formulated. Diet 1 was a control diet based on fish meal, soy protein concentrate, and wheat gluten meal. Diets 2, 3, and 4 were formulated so that P. variotii replaced 5, 10, and 20% of the CP content of the diets, respectively. Results showed that dietary inclusion of P. variotii was associated with changes in physical pellet quality, including significant linear (p < 0.0001) and/or quadratic (p < 0.01) decreases in pellet length, width, expansion, and durability. Conversely, significant linear (p < 0.0001) and quadratic (p < 0.025) increases in water activity, bulk density, and water stability index were associated with increasing replacement of the CP content of the diets with P. variotii. There were no significant differences in weight gain, growth rate or feed intake among fish fed the experimental diets (p > 0.05), but a significant linear improvement (p < 0.025) in feed conversion ratio (FCR) with increasing dietary inclusion of P. variotii was observed. The apparent digestibility coefficients for CP and gross energy of the experimental diets decreased linearly (p < 0.01) with increasing dietary levels of P. variotii. Similar results were observed for the essential and non-essential amino acids. Despite reduced digestibility, linear increases in nitrogen (p < 0.01), energy (p < 0.025), and mineral (p < 0.001) retention efficiencies were associated with increasing dietary levels of P. variotii. In the distal intestine, dietary inclusion of P. variotii resulted in significant (p < 0.05) upregulation of several cytokines (ifng, il10, and tgfb), effector molecules (inos, arg1, sod), and transcription factors (irf4), suggesting both activation and control of the inflammatory response in this key mucosa-associated lymphoid tissue. Overall, replacement of 20% of the CP content of the diet with P. variotii reduced digestibility of the experimental diets, but did not affect growth performance and improved FCR and nutrient utilization efficiency of salmon. Thus, P. variotii is a promising ingredient with potential for use in salmonid aquafeeds to improve fish performance and health.
•P. variotii can replace 20% of the crude protein content in diets for Atlantic salmon.•Dietary inclusion of P. variotii significantly improved feed conversion ratio.•P. variotii was associated with increases in nutrient utilization efficiency.•P. variotti was associated with improved gut health.•P. variotii exhibited immuno-modulatory capacity in the distal intestine of salmon.
The demand to develop protein production systems that are both economically and scientifically viable is reflected in the global scenario, where filamentous fungi, due to their interesting ...characteristics such as the high capacity to secrete proteins into the culture medium, growth in relatively simple substrates and robust post-translational machinery, among others, are presented as promising alternatives for the creation and establishment of these systems. Currently, these organisms produce a wide range of proteins, such as glycosidases, lipases, and proteases, for example. Scientific and technological development has increasingly allowed the evolution of molecular biology techniques that facilitate the genetic modification of organisms, thus, stimulating the establishment of new protein production systems. Amongst these techniques, it is possible to highlight the CRISPR/Cas system, a relatively simple, low-cost, and high-efficient tool for genetic modifications. Filamentous fungi, organisms widely used for protein production, have been used in a relatively low number of studies related to the production of (hemi-)cellulases using the CRISPR/Cas system as a genomic editing tool. (Hemi-)cellulases, enzymes that catalyze the breakdown of saccharides, are a class of enzymes that are highly researched and applied in several biotechnological areas in order to obtain a wide range of value-added bioproducts, such as bioethanol, for example. In this context, this review aims to illustrate the scenario of the application of the CRISPR/Cas technique for the production of (hemi-)cellulases, highlighting the main studies to date and the perspectives of a market that tends to grow exponentially in the coming years.
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•Protein production global market has grown exponentially in recent years.•CRISPR/Cas technology paves the way to further expand the bioengineering protein market.•Hitherto, only a few studies have reported the application of CRISPR/Cas to produce (hemi)-cellulases in filamentous fungi.
Microbial cells do not live in isolation in their environment, but rather they communicate with each other using chemical signals. This sophisticated mode of cell-to-cell signalling, known as quorum ...sensing, was first discovered in bacteria, and coordinates the behaviour of microbial population behaviour in a cell-density-dependent manner. More recently, these mechanisms have been described in eukaryotes, particularly in fungi, where they regulate processes such as pathogenesis, morphological differentiation, secondary metabolite production and biofilm formation. In this manuscript, we review the information available to date on these processes in yeast, dimorphic fungi and filamentous fungi. We analyse the diverse chemical 'languages' used by different groups of fungi, their possible cross-talk and interkingdom interactions with other organisms. We discuss the existence of these mechanisms in multicellular organisms, the ecophysiological role of QS in fungal colonisation and the potential applications of these mechanisms in biotechnology and pathogenesis.
Rapid and accurate mold identification is critical for guiding therapy for mold infections. MALDI-TOF MS has been widely adopted for bacterial and yeast identification; however, few clinical ...laboratories have applied this technology for routine mold identification due to limited database availability and lack of standardized processes. Here, we evaluated the versatility of the NIH Mold Database in a multicenter evaluation.
The NIH Mold Database was evaluated by eight US academic centers using a solid media extraction method and a challenge set of 80 clinical mold isolates. Multiple instrument parameters important for spectra optimization were evaluated, leading to the development of two specialized acquisition programs (NIH method and the Alternate-B method).
A wide range in performance (33-77%) was initially observed across the eight centers when routine spectral acquisition parameters were applied. Use of the NIH or the Alternate-B specialized acquisition programs, which are different than those used routinely for bacterial and yeast spectral acquisition (MBT_AutoX), in combination with optimized instrument maintenance, improved performance, illustrating that acquisition parameters may be one of the key limiting variable in achieving successful performance.
Successful mold identification using the NIH Database for MALDI-TOF MS on Biotyper systems was demonstrated across multiple institutions for the first time following identification of critical program parameters combined with instrument optimization. This significantly advances our potential to implement MALDI-TOF MS for mold identification across many institutions. Because instrument variability is inevitable, development of an instrument performance standard specific for mold spectral acquisition is suggested to improve reproducibility across instruments.
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.
In marine ecosystems, macroalgae are the habitat for several microorganisms, fungi being among them. In the Antarctic benthic coastal ecosystem, macroalgae play a key role in organic matter cycling. ...In this study, 13 different macroalgae from Potter Cove and surrounding areas were sampled and 48 fungal isolates were obtained from six species, four Rhodophyta Ballia callitricha, Gigartina skottsbergii, Neuroglossum delesseriae and Palmaria decipiens, and two Phaeophyceae: Adenocystis utricularis and Ascoseira mirabilis. Fungal isolates mostly belonged to the Ascomycota phylum (Antarctomyces, Cadophora, Cladosporium, Penicillium, Phialocephala, and Pseudogymnoascus) and only one to the phylum Mucoromycota. Two of the isolates could not be identified to genus level, implying that Antarctica is a source of probable novel fungal taxa with enormous bioprospecting and biotechnological potential. 73% of the fungal isolates were moderate eurypsychrophilic (they grew at 5–25 °C), 12.5% were eurypsychrophilic and grew in the whole range, 12.5% of the isolates were narrow eurypsychrophilic (growth at 15–25 °C), and Mucoromycota AUe4 was classified as stenopsychrophilic as it grew at 5–15 °C. Organic extracts of seven macroalgae from which no fungal growth was obtained (three red algae Georgiella confluens, Gymnogongrus turquetii, Plocamium cartlagineum, and four brown algae Desmarestia anceps, D. Antarctica, Desmarestia menziesii, Himantothallus grandifolius) were tested against representative fungi of the genera isolated in this work. All extracts presented fungal inhibition, those from Plocamium cartilagineum and G. turquetii showed the best results, and for most of these macroalgae, this represents the first report of antifungal activity and constitute a promising source of compounds for future evaluation.
•Several Antarctic macroalgae are habitat for marine and cosmopolitan fungi.•Some macroalgae present antifungal activities with biotechnological potential.•Isolated fungi showed a different spectrum of growth temperatures.
Seafood has an important role to play to achieve a sustainable food system that provides healthy food to a growing world population. Future seafood production will be increasingly reliant on ...aquaculture where feed innovation is essential to reduce environmental impacts and minimize feed and food competition. This study aimed to investigate whether a novel single cell protein feed ingredient based on Paecilomyces variotii grown on a side stream from the forest industry could improve environmental sustainability of farmed rainbow trout (Oncorhynchus mykiss) by replacing the soy protein concentrate used today. A Life Cycle Assessment including commonly addressed impacts but also the rarely assessed biodiversity impacts was performed. Furthermore, feeding trials were included for potential effects on fish growth, i.e., an assessment of the environmental impacts for the functional unit 'kg feed required to produce 1 kg live-weight rainbow trout'. Results showed that the best experimental diet containing P. variotii performed 16-73 % better than the control diet containing soy protein concentrate in all impact categories except for energy demand (21 % higher impact). The largest environmental benefits from replacing soy protein with P. variotii in rainbow trout diets was a 73 % reduction of impact on biodiversity and halved greenhouse gas emissions. The findings have high relevance for the aquaculture industry as the production scale and feed composition was comparable to commercial operations and because the effect on fish growth from inclusion of the novel ingredient in a complete diet was evaluated. The results on biodiversity loss from land use change and exploitation through fishing suggest that fishery can dominate impacts and exclusion thereof can greatly underestimate biodiversity impact. Finally, a novel feed ingredient grown on side streams from the forest industry has potential to add to food security through decreasing the dependence on increasingly scarce agricultural land resources.
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•Interactome represents a whole set of interactions during lignocellulose breakdown.•Fungus-substrate interactome drives enzyme expression.•Enzyme-enzyme interactome involves ...synergism and LMW molecules exchanging.•Interactome can be driven to enhance biomass valorization.•Interactome can unlock the full potential of lignocellulosic components.
Biomass represents an abundant and inexpensive source of sugars and aromatic compounds that can be used as raw materials for conversion into value-added bioproducts. Filamentous fungi are sources of plant cell wall degrading enzymes in nature. Understanding the interactions between enzymes is crucial for optimizing biomass degradation processes. Herein, the concept of the interactome is presented as a holistic approach that depicts the interactions among enzymes, substrates, metabolites, and inhibitors. The interactome encompasses several stages of biomass degradation, starting with the sensing of the substrate and the subsequent synthesis of hydrolytic and oxidative enzymes (fungus-substrate interaction). Enzyme-enzyme interactions are exemplified in the complex processes of lignocellulosic biomass degradation. The enzyme-substrate-metabolite-inhibitor interaction also provides a better understanding of biomass conversion, allowing bioproduct production from recalcitrant agro-industrial residues, thus bringing greater value to residual biomass. Finally, technological applications are presented for optimizing the interactome at various levels.
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