Search Results (181)

Cultivable fungi are important components of freshwater ecosystems, yet their diversity in Amazonian aquatic environments remains poorly explored. This study evaluated cultivable fungal communities associated with water, sediment, and submerged wood in the Negro, Solimões, and Tapajós Rivers, representing the major black-, white-, and clear-water systems of the Amazon basin, respectively. Samples were collected along 25 m transects, fungi were isolated on potato dextrose agar, grouped into morphotypes, and identified morphologically. Diversity was assessed using richness, Shannon, Simpson, Pielou, and Sørensen indices. Overall, 130 isolates and 75 morphotypes were recorded, with a predominance of morphotypes assigned to filamentous Ascomycota and widespread occurrence of Aspergillus and Penicillium. Solid substrates yielded greater fungal abundance and richness than water samples. Submerged wood from the Negro River showed the highest overall cultivable fungal abundance, whereas the Tapajós River showed the highest diversity in water and sediment and the highest richness in both substrates. The Solimões River showed stronger dominance by a limited number of morphotypes, particularly in submerged wood. Sørensen similarity values indicated low compositional overlap among rivers, especially for submerged wood communities, suggesting apparent differentiation among river-associated cultivable fungal assemblages. Together, these exploratory results suggest that substrate type, hydrochemical differences, and potential temporal effects may be associated with the structure of cultivable fungal communities in Amazonian rivers. Full article

The increasing demand for safer and environmentally sustainable products has intensified the search for natural alternatives to synthetic dyes. Filamentous fungi are promising sources of natural pigments due to their metabolic diversity and the feasibility of large-scale production. In this study, filamentous fungi isolated from Amazonian freshwater environments were evaluated for their potential to produce natural pigment-associated metabolites under different nutritional conditions. Forty-five fungal isolates were screened in solid media and subsequently cultivated in submerged fermentation using three media: potato dextrose broth supplemented with yeast extract (BD + YE); malt extract broth (ME); and yeast extract–sucrose broth supplemented with magnesium sulfate (YES). Among the 39 pigment-producing isolates, seven were selected for further investigation. Sucrose favored the highest absorbance values of pigment extracts, particularly for isolates identified as Talaromyces amestolkiae. In addition, the extract of T. amestolkiae TA10P5-3 exhibited the highest absorbance value (6.83 abs. units at 400 nm) when cultivated in YES medium, indicating stronger chromophore-associated spectral signals. This extract also showed antimicrobial activity against Pseudomonas aeruginosa (625 μg/mL), Staphylococcus epidermidis (312 μg/mL), and Candida tropicalis (625 μg/mL). Finally, the TA10P5-3 extract presented high total phenolic content (246.30 mg GAE/g) and antioxidant activity (EC₅₀ = 5470 μg/mL). These findings highlight Amazonian freshwater fungi as promising sources of natural pigments with potential industrial applications. Full article

Natural antioxidants are essential for protecting the body against oxidative stress and exhibit a wide range of biological activities. In this context, forty extracts derived from ten submerged cultivated mushroom species were analyzed for their mycochemical composition, antioxidant capacity, and cytotoxic effects against MCF7 breast cancer cells. Qualitative and quantitative screening revealed that, among the detected classes of bioactive compounds, the extracts were predominantly enriched in flavonoids, terpenoids, and phenolic constituents. Considerable variation was observed in the levels of total phenolics, flavonoids, and ascorbic acid among different species and solvent extracts. The highest total phenolic contents were detected in ethanol and ethyl acetate extracts of G. frondosa (110.0 ± 6.4, 227.6 ± 14.2, and 160.5 ± 5.3 mg GAE/g), while the water extract of F. velutipes also exhibited elevated phenolic levels (119.2 ± 6.5 mg GAE/g). Flavonoid concentrations ranged from 102.5 ± 10.5 to 359.9 ± 2.5 mg QE/g in biomass and culture liquid extracts obtained with organic solvents. Ascorbic acid content was generally highest in ethyl acetate culture liquid extracts, suggesting solvent-dependent enrichment of antioxidant metabolites. Free radical scavenging activity increased in a concentration-dependent manner, reaching inhibition values more than 90% at 20 mg/mL in all tested mushrooms. Cytotoxicity assays demonstrated that extract type, solvent, and incubation time strongly influenced the inhibition of MCF7 cell viability. Ethyl acetate extracts from H. erinaceus, P. ostreatus, T. versicolor, and T. pubescens exhibited the strongest cytotoxic effects, reducing cell viability by up to 70% at higher concentrations. The results demonstrate that mushroom extracts, particularly ethyl acetate extracts, possess significant antioxidant and cytotoxic activities. These findings highlight their potential as promising natural sources of medicinal bioactive compounds for antioxidant and anticancer applications. Full article

Thraustochytrids are promising alternatives for the production of docosahexaenoic acid (DHA; C22:6 n-3), a long-chain polyunsaturated fatty acid with health benefits. For practical application of this oleaginous microorganism, an efficient cultivation method to enhance DHA production is required, which relies on several factors that support cell growth, lipid accumulation, and lipid turnover. In this study, the robust submerged fermentation of an acid- and high-temperature-tolerant strain of Aurantiochytrium limacinum was investigated. Under controlled temperature and acidic conditions (pH 4.5 and 30 °C), glucose and peptone were the best carbon and nitrogen sources for enhancing biomass and DHA production, respectively, with a glucose concentration of 60 g/L and a C/N ratio of 24 being optimal for DHA production. Applying an aeration rate of 2 vvm and an agitation speed of 300 rpm using a combination of a ring sparger and pitch-blade impeller in a stirred-tank bioreactor improved DHA production using intermittent fed-batch fermentation. The highest DHA titer was obtained at 3.01 g/L, and the DHA content in biomass was 10.69% (w/w) after intermittent feeding of cassava starch hydrolysate as the substrate. Full article

Antimicrobial metabolite production in basidiomycetes varies by strain and growing conditions. This study compared the antimicrobial activity of extracts from nine fungal strains at both their vegetative and reproductive stages. Wild-growing fungal fruiting bodies were collected and identified through both morphological characterization and molecular sequencing. Extracts from fruiting bodies, mycelia, and culture liquids were tested using the agar well diffusion method and by determining the minimum inhibitory concentration (MIC). Analysis revealed that the highest antimicrobial activity was associated with culture liquid extracts. Antimicrobial properties were detected in the submerged mycelium extracts of only two strains: Stereum hirsutum 1 and Flammulina rossica 16. For fruiting bodies, activity was restricted to extracts of strains from the genus Fomitopsis. The strain S. hirsutum 1 was determined to be the most effective producer of antibacterial compounds. The highest activity was exhibited by the S. hirsutum 1 culture liquid extract, with an MIC of 320 µg/mL against clinical Gram-positive strains (Staphylococcus aureus, S. epidermidis, S. haemolyticus, vancomycin-resistant Enterococcus faecium) and Gram-negative Proteus vulgaris. The studied strains demonstrated higher production of antimicrobial metabolites during vegetative growth, with the active compounds being primarily extracellular. Submerged cultivation of basidiomycetes offers an efficient method for obtaining antimicrobial metabolites, permitting their subsequent isolation, physicochemical characterization, and biomedical evaluation. Full article

Hypocrellin A (HA) represents a pharmaceutically important perylenequinone photosensitizer produced by Shiraia bambusicola. However, submerged fermentation remains constrained by filamentous morphological characteristics and inherent mass transfer limitations. Although microparticle-enhanced cultivation (MPEC) has demonstrated efficacy in filamentous fungal systems, the molecular mechanisms by which physical cues, such as microparticle-induced shear stress, reprogram fungal metabolism remain largely unexplored. This work systematically optimizes SiO₂-based MPEC parameters for S. bambusicola GDMCC 60438, including particle dimensions, temporal addition protocols, and solid loading. Mechanistic investigations integrated pellet morphology analysis, membrane lipid composition, intracellular redox status, energy/precursor markers, and RNA-seq transcriptomic profiling with qRT-PCR validation. Under optimized conditions (10% w/v SiO₂, 30 mesh, added at 6 h), HA yield reached 41.76 ± 5.02 mg/L, representing a 3.65-fold increase over controls. MPEC shifted morphology toward smaller, more porous pellets with denser internal structure, accompanied by increased membrane fluidity (unsaturated/saturated fatty acid ratio from 1.54 to 2.63), elevated ROS levels with antioxidant enzyme activation, and enhanced acetyl-CoA and ATP accumulation. Transcriptomic analysis identified 206 differentially expressed genes enriched in oxidative phosphorylation, carbon metabolism, and stress responses, with upregulation of PKS-related biosynthetic genes and major facilitator superfamily transporters. This work establishes an integrated mechanistic framework linking particle-induced morphological changes to metabolic reprogramming through oxidative stress and subsequent transcriptional activation of the HA biosynthetic pathway, providing rational design principles for MPEC strategies in filamentous fungi. Full article

In this work, a food-compatible bioprocess was evaluated for the production of yeast single-cell protein from mezcal agave bagasse. Bagasse was enzymatically hydrolyzed at 10% (w/v) solids (pH 4.8, 50 °C, 24 h) using commercial enzymes. The resulting liquid was clarified by activated charcoal adsorption and filtration to obtain a hydrolysate suitable for submerged fermentation. Enzymatic hydrolysis released reducing sugars in the range of 11–17 g/L. Saccharomyces cerevisiae was cultivated on the clarified hydrolysate under submerged conditions using both flask-scale and 2 L stirred-tank bioreactor experiments. Trials were performed at flask scale with initial sugars at 8, 17, and 50 g/L, and at 2 L stirred-tank bioreactor scale with initial sugars at 20.68 g/L (R1) and 16.30 (R2) g/L. At the flask scale, final biomass concentrations increased with initial sugar level. Values reached 6.18 ± 0.27, 8.02 ± 0.55, and 9.28 ± 0.10 g/L, while crude protein remained below 10% (3.40 ± 0.15 to 8.69 ± 0.09 g/100 g dry weight). In contrast, bioreactor cultivation resulted in higher protein enrichment, with protein contents over 40% under both oxygen regimes (41.71 ± 0.47 to 45.80 ± 0.43 g/100 g dry weight). Overall, the findings support enzymatic hydrolysis coupled with controlled submerged fermentation as a scalable approach for valorizing agave bagasse into protein-enriched yeast biomass. Full article

The present study investigates the potential of olive mill wastewater (OMW), supplemented with expired commercial glucose syrup, as a sustainable substrate for the submerged cultivation of Tuber spp. wild mushrooms. OMW contains considerable quantities of phenolic compounds, making it both a challenging pollutant and a promising nutrient source. To assess fungal performance under increasing phenolic stress, culture media were prepared with varying OMW concentrations (0–75% v/v on agar; 0–50% v/v in liquid media), while glucose was adjusted to ~30 g/L using expired glucose syrup. A sequential experimental approach was followed, beginning with Petri dish screenings on substrate/strain selection (measuring the mycelial growth rate; Kr, mm/day), progressing to 25-day shake flask fermentations and subsequently scaling up the most promising strain (Tuber mesentericum) in a controlled stirred-tank bioreactor. Throughout cultivation, substrate consumption (glucose, phenolics), pH evolution and decolorization were evaluated, while the resulting biomass was analyzed for polysaccharides, β-glucans, proteins, lipids, fatty acids, antioxidants, phenolic acids and triterpenoids content. Results showed that increasing OMW concentration enhanced tolerance and metabolic activity in selected Tuber species, with T. mesentericum exhibiting the highest resilience and achieving comparable or higher biomass yields in OMW-based media than in glucose (control). Phenolic removal exceeded 60% in flasks and 50% in the bioreactor, confirming simultaneous bioremediation capacity. Bioreactor cultivation demonstrated efficient substrate utilization and biomass production, while OMW-grown biomass presented high lipid content, enriched with unsaturated fatty acids, high β-glucan levels and increased antioxidant and phenolic profiles. Overall, this study demonstrates that OMW (supplemented with expired glucose syrup) can serve as a cost-effective and environmentally beneficial substrate for Tuber biomass production with dietary and antioxidant properties, offering an alternative source to mushroom carposomes, as well as supporting the circular bioeconomy strategies within olive oil processing industries. Full article

Xylaria karsticola NBIMCC 9097 is a recently described and rare fungicolous species originating from Bulgaria. Understanding its growth behavior and bioactive potential is essential for evaluating its biotechnological and pharmaceutical relevance. In the presented study, we model the in vitro growth kinetics of X. karsticola mycelium under submerged cultivation and assess its antimicrobial activity. Optimization of MCM and MYB media markedly increased biomass yields to 20.11 and 23.25 g/dm³, respectively, compared with non-optimized media (9.9 ± 0.21 and 10.8 ± 0.28 g/dm³). The maximum specific growth rate was higher in the MCM (0.803 ± 0.004 h⁻¹) in comparison with the MYB medium (0.711 ± 0.003 h⁻¹); however, the MYB medium supported greater biomass accumulation and more efficient substrate utilization, reflected by a higher utilization coefficient (0.9900 ± 0.001 versus 0.9644 ± 0.005). The antimicrobial activity was evaluated using agar disk diffusion and minimum inhibitory concentration assays against Gram-positive and Gram-negative bacteria and yeasts. Hexane and ethyl acetate extracts were most effective against Pseudomonas aeruginosa ATCC 9027 (MIC 0.067 and 0.059 mg/cm³), while notable anti-yeast activity was observed, particularly against Wickerhamomyces anomalus, Saccharomycodes ludwigii, and Pichia membranifaciens. The lowest MIC (0.02 mg/cm³) was recorded for the water biomass extract against S. ludwigii indicating potent antimicrobial activity against the tested microorganism. These findings identify X. karsticola as a potential source of antimicrobial metabolites and provide a strong motivation for comprehensive metabolomic profiling and systematic optimization of its cultivation. Full article

This pioneering study investigates bacteria isolated from marine pelagic fish, fishing vessels, and gear surfaces, focusing on the variability in biofilm formation across different substrates, media, and cultivation conditions. Bacteria from fish intestines, skin, and gills, including spoilage organisms and potential fish and human pathogens, can contaminate vessel surfaces, gear, and containers and may act as microbial reservoirs and transmission vectors. In this study, biofilm formation was evaluated at air–liquid interfaces and on submerged plastic, metal, and glass surfaces under various incubation temperatures and media. Vibrio spp. were isolated both from fishing nets and fish gills, particularly Vibrio alginolyticus, V. gigantis, and V. pelagius. Although V. harveyi was examined as a representative vibrio, it did not form a biofilm on polypropylene. Photobacterium damselae subsp. damselae, Pseudomonas fragi, P. gessardii, Psychrobacter spp., and Rothia endophytica showed a strong affinity for stainless steel. Overall adhesion regardless of media type was highest for P. gessardii, followed by P. damselae and Aeromonas veronii, which adhered strongly to steel, glass, and polypropylene; however, only P. gessardii also adhered well to polystyrene, an important finding because these are known fish and human pathogens. These results highlight species-dependent biofilm triggers and their substantial variability and provide guidance for standardized marine biofilm protocols. Full article

The present study explores the treatment and valorization of carbon-rich, low-cost waste streams—sugar beet molasses, expired rice, and wheat cereal hydrolysates—as substrates for submerged shake-flask cultures of edible ascomycetes (Morchella elata AMRL 63, Tuber aestivum AMRL 364) and basidiomycetes (Lentinula edodes AMRL 126, Agaricus bisporus AMRL 209) within a circular bioeconomy framework. Cultures were conducted under different C/N ratios (20 and 50) with or without the addition of olive oil or its emulsion. Among the tested species, the ascomycetes M. elata AMRL 63 and T. aestivum AMRL 364 outperformed the basidiomycetes in biomass production and substrate utilization. Supplementation with olive oil or its emulsion enhanced mycelial growth and lipid accumulation, while a higher C/N ratio (50) favored exopolysaccharide (EPS) synthesis. Lipid profiles were dominated by oleic (Δ⁹C18:1) and linoleic (Δ^9,12C18:2) acids, with greater unsaturation observed in C/N = 20 cultures. Volatile analysis revealed species-specific aroma signatures, including characteristic truffle and morel compounds. The results underscore the feasibility of using waste streams for sustainable mushroom cultivation. Full article

Obtaining enzymes through bioconversion depends on a complex relationship between the microorganisms and the biomass used. Here, we evaluate xylanase production by diverse actinobacterial species, cultivated using xylan as the sole carbon source and complex media containing sorghum as the substrate. Fifty-three actinobacteria were tested for xylanase production in a solid medium. Seventeen strains produced xylanase and were tested for their ability to produce xylanase, total cellulases (filter paper activity, FPase), and endoglycanase in submerged culture using a defined liquid medium. The best xylanase-producing species was Streptomyces capoamus, yielding 24 IU·mL⁻¹. For FPase, Streptomyces sp. showed the highest yield (1.12 IU·mL⁻¹); for endoglycanase, the best producer was Streptomyces ossamyceticus (0.99 IU·mL⁻¹). When sweet sorghum was used alone, S. curacoi, S. ossamyceticus, and S. capoamus showed xylanase activities of 4.5 IU·mL⁻¹, 4.4 IU·mL⁻¹, and 0.8 IU·mL⁻¹, respectively. However, FPase activity was not detected under the assay conditions. The results showed that there is an intraspecific difference in xylanase, endoglucanase, and FPase production by actinobacteria, with the species S. curacoi, S. ossamyceticus, and S. capoamus able to use sorghum as a carbon source, demonstrating biotechnological potential. Full article

Rice paddy fields (referred to below as rice fields) are important not only for food production, but also as habitats for various species. The Nagoya Daruma Pond Frog (Pelophylax porosus brevipodus) is an endangered frog species endemic to Japan, mainly living in and around rice field areas. In July 2018, heavy rainfall caused severe flooding in Mabi Town of Okayama Prefecture, western Japan, submerging numerous rice fields and affecting local frog populations, including P. porosus brevipodus. To clarify whether the population structure of P. porosus brevipodus changed following the flood disaster in the rice fields of Mabi Town, we conducted quantitative field surveys in a rice fallow field in mid-October before (2017) and after (2018, 2020–2022, excluding 2019) the flood. The number of frogs declined sharply after the 2018 flood, reaching only a few individuals by 2020, but showed a substantial recovery in 2021 following the resumption of rice cultivation, although numbers decreased again in 2022. This recovery, despite fluctuations, indicates that habitat restoration through rice farming played a key role in enabling the population to rebound. Our findings underscore the importance of maintaining and restoring rice field environments after natural disasters for the survival and long-term recovery of P. porosus brevipodus. Full article

Fermented forest litter (FFL) is a biofertilizer obtained by anaerobic fermentation of forest litter combined with agricultural by-products. Its production involves an initial one-month solid-state fermentation of oak litter mixed with whey, molasses and wheat bran, followed by a one-week submerged fermentation-called the “activation” phase-during which the solid FFL is fermented with sugarcane molasses diluted in water. This study aimed to evaluate the effects storage duration (6, 18 and 30 months), and temperature (ambient and 29 °C) on the activation phase. For this purpose, pH, sugar consumption and metabolite production dynamics were monitored. Under all experimental conditions, the pH dropped to values close to 3.5, sucrose was rapidly hydrolyzed, and glucose was preferentially consumed over fructose. Fructose was metabolized only after glucose was depleted, suggesting the involvement of fructophilic microorganisms. The time-course evolution of lactic acid (LA) concentration was adequately fitted by the Gompertz model (R² > 0.970). The highest LA_max concentration (6.30 g/L) and production rate (2.16 g/L·d) were obtained with FFL stored for 6 months. Acetic acid (AA) and ethanol were also detected reaching maxima values of 1.19 g/L and 0.96 g/L, respectively. Their profiles varied depending on the experimental conditions. Notably, the AA/LA ratio increased with the age of the FFL. Overall, sugar consumption and metabolite production were significantly slower at ambient temperature, than at 29 °C. These results contribute to a better understanding of the metabolic dynamics during FFL activation and highlight key parameters that should be considered to optimize future biofertilizer production processes. Full article

The worldwide population is anticipated to reach 10.12 billion by the year 2100, thereby amplifying the necessity for sustainable agricultural methodologies to secure food availability while reducing ecological consequences. Conventional synthetic pesticides, while capable of increasing crop yields by as much as 50%, present considerable hazards such as toxicity, the emergence of resistance, and environmental pollution. This review examines biopesticides, originating from microbial (e.g., Bacillus thuringiensis, Trichoderma spp.), plant, or animal sources, as environmentally sustainable alternatives which address pest control through mechanisms including antibiosis, hyperparasitism, and competition. Biopesticides provide advantages such as biodegradability, minimal toxicity to non-target organisms, and a lower likelihood of resistance development. The global market for biopesticides is projected to be valued between USD 8 and 10 billion by 2025, accounting for 3–4% of the overall pesticide sector, and is expected to grow at a compound annual growth rate (CAGR) of 12–16%. To mitigate production costs, agro-industrial byproducts such as rice husk and starch wastewater can be utilized as economical substrates in both solid-state and submerged fermentation processes, which may lead to a reduction in expenses ranging from 35% to 59%. Strategies for process intensification, such as the implementation of intensified bioreactors, continuous cultivation methods, and artificial intelligence (AI)-driven monitoring systems, significantly improve the upstream stages (including strain development and fermentation), downstream processes (such as purification and drying), and formulation phases. These advancements result in enhanced productivity, reduced energy consumption, and greater product stability. Patent activity, exemplified by 2371 documents from 1982 to 2021, highlights advancements in formulations and microbial strains. The integration of circular economy principles in biopesticide production through process intensification enhances the safety, quality, and sustainability of food systems. Projections suggest that by the 2040s to 2050s, biopesticides may achieve market parity with synthetic alternatives. Obstacles encompass the alignment of regulations and the ability to scale in order to completely achieve these benefits. Full article