Search Results (70)

Almonds are an essential crop for the economy of California. However, mold and mycotoxin contamination of this commodity has a serious impact on food safety and international trade. The contamination levels of molds and the aflatoxigenic potential of Aspergillus section Flavi isolates were studied on almonds collected at a processing plant in California. The mean total fungal count for 80 samples was 1.0 × 10⁴ CFU/g, while 62 samples (77.5%) had a total mold count less than 1.0 × 10⁴ CFU/g. The most common fungal contaminants were Aspergillus section Nigri (100% of samples), followed by Penicillium (57.5%) and Cladosporium (52.5%) species. Rhizopus, Fusarium and Alternaria spp. were less frequent. A total of 26 A. section Flavi strains were identified, with most strains (23) belonging to the L morphotype of A. flavus. In addition, two S morphotypes of A. flavus, and one A. tamarii strain were observed. Other Aspergillus species, including A. terreus and A. ochraceus were rare. High Performance Liquid Chromatography (HPLC) analysis revealed that 9 out of 13 isolated A. flavus strains produced aflatoxin B₁ (AFB₁) on yeast extract sucrose media. The highest levels of AFB₁ were produced by two A. flavus isolates belonging to the S morphotype (78 and 260 µg/kg). Increasing temperatures and drought conditions may change the population dynamics of toxigenic mold strains on almonds, emphasizing the need to continue monitoring these fungal populations. Full article

Rice is one of the most important staple foods for the human population, necessitating continuous monitoring for mycotoxin risk in particular in the sub-tropical area, such as India. In the present study, a total of eighty-one samples comprising brown (n = 36) and polished (white) rice (n = 45) intended for direct human consumption were collected from markets across various districts of Tamil Nadu, India, and analysed for ochratoxin A (OTA) and fungal contamination. Aspergillus ochraceus, an ochratoxigenic fungus belonging to Aspergillus section Circumdati, exhibits optimal growth and OTA production at temperatures ranging from 25 °C to 30 °C. Among the fungal isolates, Aspergillus niger and A. ochraceus were the most prevalent, occurring in 50 out of 81 samples (62%). A. ochraceus demonstrated a significantly higher OTA-producing capacity compared to A. niger, with an OTA concentration range of 12.3–196.8 µg/kg and 0.2–2.8 µg/kg. Chemical analysis of fifty fungal-contaminated market rice samples revealed that 76% (38/50) were contaminated with OTA. Further, detectable levels of OTA were observed in 83% of brown rice and 69% of polished rice samples, with the highest frequency falling within the range of 1–<3 µg/kg. However, none of the tested rice samples exceeded the acceptable OTA threshold set by the Food Safety and Standards Authority of India (FSSAI) (20 µg/kg), with all concentrations falling below the national regulatory limit. This study represents further insight into OTA exposure in rice, with greater concern regarding brown rice than white rice, and emphasizes the necessity of implementing sound and safe storage practices, effective management strategies, and continuous monitoring programs to prevent OTA contamination throughout the Indian rice supply chain. Full article

Several species of Fusarium are reported to infect inflorescences of high-THC-containing cannabis (Cannabis sativa L.) plants grown in greenhouses in Canada. These include F. graminearum, F. sporotrichiodes, F. proliferatum, and, to a lesser extent, F. oxysporum and F. solani. The greatest concern surrounding the infection of cannabis by these Fusarium species, which cause symptoms of bud rot, is the potential for the accumulation of mycotoxins that may go undetected. In the present study, both naturally infected and artificially infected inflorescence tissues were tested for the presence of fungal-derived toxins using HPLC-MS/MS analysis. Naturally infected cannabis tissues were confirmed to be infected by both F. avenaceum and F. graminearum using PCR. Pure cultures of these two species and F. sporotrichiodes were inoculated onto detached inflorescences of two cannabis genotypes, and after 7 days, they were dried and assayed for mycotoxin presence. In these assays, all Fusarium species grew prolifically over the tissue surface. Tissues infected by F. graminearum contained 3-acetyl DON, DON, and zearalenone in the ranges of 0.13–0.40, 1.18–1.91, and 31.8 to 56.2 μg/g, respectively, depending on the cannabis genotype. In F. sporotrichiodes-infected samples, HT2 and T2 mycotoxins were present at 13.9 and 10.9 μg/g in one genotype and were lower in the other. In F. avenaceum-inoculated tissues, the mycotoxins enniatin A, enniatin A1, enniatin B, and enniatin B1 were produced at varying concentrations, depending on the isolate and cannabis genotype. Unexpectedly, these tissues also contained detectable levels of 3-acetyl DON, DON, and zearalenone, which was attributed to apre-existing natural infection by F. graminearum that was confirmed by RT-qPCR. Beauvericin was detected in tissues infected by F. avenaceum and F. sporotrichiodes, but not by F. graminearum. Naturally infected, dried inflorescences from which F. avenaceum was recovered contained beauvericin, enniatin A1, enniatin B, and enniatin B1 as expected. Uninoculated cannabis inflorescences were free of mycotoxins except for culmorin at 0.348 μg/g, reflecting pre-existing infection by F. graminearum. The mycotoxin levels were markedly different between the two cannabis genotypes, despite comparable mycelial colonization. Tall fescue plants growing in the vicinity of the greenhouse were shown to harbor F. avenaceum and F. graminearum, suggesting a likely external source of inoculum. Isolates of both species from tall fescue produced mycotoxins when inoculated onto cannabis inflorescences. These findings demonstrate that infection by F. graminearum and F. avenaceum, either from artificial inoculation or natural inoculum originating from tall fescue plants, can lead to mycotoxin accumulation in cannabis inflorescences. However, extensive mycelial colonization following prolonged incubation of infected tissues under high humidity conditions is required. Inoculations with Penicillium citrinum and Aspergillus ochraceus under these conditions produced no detectable mycotoxins. The mycotoxins alternariol and tentoxin were detected in several inflorescence samples, likely as a result of natural infection by Alternaria spp. Fusarium avenaceum is reported to infect cannabis inflorescences for the first time and produces mycotoxins in diseased tissues. Full article

The major objective was to investigate the protective capabilities of endophytic bacterial strains isolated from a number of medicinal plant species towards Aspergillus spp. secured from the internal tissues of fungi-infected peanuts. Among 32 fungal isolates surveyed for mycotoxin production in various culture media (PDA, RBCA, YES, CA), 10 isolates qualitatively producing AFB₁, besides 10 OTA-producers, were assayed by HPLC for quantitative toxin production. Aspergillus spp. isolate Be 13 produced an extraordinary quantity of 1859.18 μg mL⁻¹ AFB₁, against the lowest toxin level of 280.40 μg mL⁻¹ produced by the fungus isolate IS 4. The estimated amounts of OTA were considerably lower and fell in the range 0.88–6.00 μg mL⁻¹; isolate Sa 1 was superior, while isolate Be 7 seemed inferior. Based on ITS gene sequencing, the highly toxigenic Aspergillus spp. isolates Be 13 and Sa 1 matched the description of A. novoparasiticus and A. ochraceus, respectively, ochraceus, respectively, which are present in GenBank with identity exceeding 99%. According to 16S rRNA gene sequencing, these antagonists labeled Ar6, Ma27 and So34 showed the typical characteristics of Pseudomonas aeruginosa, Bacillus subtilis and Bacillus velezensis, respectively, with similarity percentages of 99–100. The plant growth-promoting activity measurements of the identified endophytes indicated the production of 16.96–80.00 μg/100 mL culture medium of IAA. Phosphate-solubilizing capacity varied among endophytes from 2.50 to 21.38 μg/100 mL. The polysaccharide production pool of bacterial strains ranged between 2.74 and 6.57 mg mL⁻¹. P. aeruginosa Ar6 and B. velezensis successfully produced HCN, but B. subtilis failed. The in vitro mycotoxin biodegradation potential of tested bacterial endophytes indicated the superiority of B. velezensis in degrading both mycotoxins (AFB₁-OTA) with average percentage of 88.7; B. subtilis ranked thereafter (85.6%). The 30-day old peanut (cv. Giza 6) seedlings grown in gnotobiotic system severely injured due to infection with AFB₁/OTA-producing fungi, an effect expressed in significant reductions in shoot and root growth traits. Simultaneous treatment with the endophytic antagonists greatly diminished the harmful impact of the pathogens; B. velezensis was the pioneer, not P. aeruginosa Ar6. In conclusion, these findings proved that several endophytic bacterial species have the potential as alternative tools to chemical fungicides for protecting agricultural commodities against mycotoxin-producing fungi. Full article

Immunotherapy is a transformative strategy in oncology, yet the development of novel immunomodulatory agents remains essential. This study explores the anti-tumor potential of a structurally unique polysaccharide isolated from an Aspergillus ochraceus (AOP), sourced from the Antarctic Weddell Sea. Using alkaline-assisted extraction and chromatographic purification, we obtained a homogeneous polysaccharide predominantly composed of galactose and mannose, with an average molecular weight of 39.67 kDa. The structure was characterized by an integrated nuclear magnetic resonance spectroscopy and mass spectrometry analysis, revealing that the AOP is composed of β (1→5)-linked galactofuranose units, with a minor substitution by α-D-mannopyranose residues via (1→2) glycosidic bonds at the C2 of the galactofuranose. Functional assays, including CCK8 and wound-healing tests, demonstrated that this polysaccharide, referred to as AOP, inhibited melanoma cell proliferation and migration in a dose-dependent manner. Additionally, the AOP activated RAW264.7 and bone marrow-derived macrophage (BMDM) cells without exhibiting significant cytotoxicity, leading to the release of inflammatory factors such as TNF-α, IL-1β, and IL-6. Mechanistically, the AOP was found to upregulate the expression of CD86 and IFN-γ, while downregulating genes like IL-4 and Arg1. These findings position the AOP as the first documented Antarctic fungal polysaccharide with macrophage-reprogramming capabilities against melanoma, offering novel molecular insights for marine-derived immunotherapeutics. Full article

Testosterone holds significant medical and economic importance, with the global market for testosterone replacement therapies valued at approximately USD 1.9 billion in 2023. This hormone is essential for the development and maintenance of male sexual characteristics as well as bone and muscle health. It plays a key role in conditions such as hypogonadism, muscle disorders, and andropause. However, the industrial production of testosterone often involves complex chemical processes that result in low yields, high costs, and environmental damage. Microbial biotransformation of steroids presents an eco-friendly alternative to traditional chemical synthesis. A knockout strain of Aspergillus nidulans deficient in steroid 11α-hydroxylase activity was developed, rendering it incapable of hydroxylating androstenedione, progesterone, and testosterone. In these strains, two newly identified CYP450 enzymes, CYP68L1 from A. nidulans and CYP68L8 from Aspergillus ochraceus, were expressed to confirm their roles as steroid 11α-hydroxylases of androstenedione, progesterone, and testosterone. The availability of these 11α-hydroxylases represents significant progress toward achieving efficient single-step steroid fermentation. Furthermore, the A. nidulans knockout strain serves as an effective model for studying the conversion of androstenedione to testosterone upon the expression of the enzyme 17β-hydroxysteroid dehydrogenase, due to its inability to hydroxylate testosterone. Full article

The present study reports the ability of a fungal isolate Aspergillus ochraceus DY1, obtained from rotten wood, to degrade alkali lignin (AL) and lignocelluloses in an efficient manner. The efficiency of degradation was monitored by measuring the percentage of decolorization and utilizing GC-MS for identifying degradation products at different time intervals (10, 20, 30, and 40 days). The optimal degradation of alkali lignin (AL) was achieved at 0.01% concentration, 25 °C, and pH 7, resulting in 63.64% degradation after 40 days of incubation. A GC-MS analysis revealed significant degradation products, including n-hexadecanoic acid, octadecane, butylated hydroxytoluene, 2,6,11-trimethyl-dodecane, dibutyl phthalate, oleic acid, 3,5-dimethoxy-phenol acetate, and 2-(phenylmethylene)- cyclohexanone. Structural changes in AL were confirmed through HSQC 2D NMR and size-exclusion chromatography, indicating depolymerization and reduced molecular weight. Furthermore, A. ochraceus DY1 demonstrated substantial biomass loss in corn stover (62.5%) and sugarcane bagasse (50%) after 7 days of solid-state fermentation. Surface morphological depletion was observed in the bio-treated corn stover through SEM and confocal microscopy, which was not seen in the untreated one. These findings underscore the potential of A. ochraceus DY1 for efficient lignin degradation, with promising applications in biofuel production, waste management in the paper and pulp industry, and the synthesis of value-added bioproducts. Full article

The growing emergence of multi-drug resistant microbial strains has kept the scientific world searching for novel bioactive compounds with specific chemical characteristics. Accordingly, researchers have started exploring the understudied metabolites from endophytes as a new source of bioactive compounds. In this context, the current study was designed to evaluate the bioactive properties of endophytic fungi from the Mokrzański forest in Wrocław, Poland that have not yet been fully researched. Forty-three endophytic fungi were isolated from twelve distinct plants. Following their cultivation, fungal extracts were separately prepared from biomass and cell-free filtrates, and their antibacterial, antifungal (against human and plant pathogens), and antioxidant properties were examined. Five promising fungi after screening were identified to possess all of these activities. These strains and their respective plant hosts were Trichoderma harzianum BUK-T (Fagus sylvatica), Aspergillus ochraceus ROB-L1 (Robinia pseudoacacia), Chaetomium cochliodes KLON-L1, Fusarium tricinctum KLON-L2 (Acer platanoides), and Penicillium chrysogenum SOS-B2 (Pinus sylvestris). Moreover, gamma irradiation at several doses (Gy) was separately applied to the fungal cultures to study their effects on the recorded activities. Finally, compounds after preparative thin-layer chromatography fractionation of the five fungal strains were identified by GC-MS. These findings suggest that the isolated endophytic fungi could serve as novel sources of bioactive metabolites with antibacterial, antifungal, and antioxidant properties, potentially paving the way for future research and the development of new bioactive compounds. Full article

Aspergillus fungi constitute a pivotal element within ecosystems, serving as both contributors of biologically active compounds and harboring the potential to cause various diseases across living organisms. The organism’s proteolytic enzyme complex, termed the degradome, acts as an intermediary in its dynamic interaction with the surrounding environment. Using techniques such as genome and transcriptome sequencing, alongside protein prediction methodologies, we identified putative extracellular peptidases within Aspergillus ochraceus VKM-F4104D. Following manual annotation procedures, a total of 11 aspartic, 2 cysteine, 2 glutamic, 21 serine, 1 threonine, and 21 metallopeptidases were attributed to the extracellular degradome of A. ochraceus VKM-F4104D. Among them are enzymes with promising applications in biotechnology, potential targets and agents for antifungal therapy, and microbial antagonism factors. Thus, additional functionalities of the extracellular degradome, extending beyond mere protein substrate digestion for nutritional purposes, were demonstrated. Full article

Nowadays, unprecedented health challenges are urging novel solutions to address antimicrobial resistance as multidrug-resistant strains of bacteria, yeasts and moulds are emerging. Such microorganisms can cause food and feed spoilage, food poisoning and even more severe diseases, resulting in human death. In order to overcome this phenomenon, it is essential to identify novel antimicrobials that are naturally occurring, biologically effective and increasingly safe for human use. The development of gemmotherapy extracts (GTEs) using plant parts such as buds and young shoots has emerged as a novel approach to treat/prevent human conditions due to their associated antidiabetic, anti-inflammatory and/or antimicrobial properties that all require careful evaluations. Seven GTEs obtained from plant species like the olive (Olea europaea L.), almond (Prunus amygdalus L.), black mulberry (Morus nigra L.), walnut (Juglans regia L.), blackberry (Rubus fruticosus L.), blackcurrant (Ribes nigrum L.) and bilberry (Vaccinium myrtillus L.) were tested for their antimicrobial efficiency via agar diffusion and microbroth dilution methods. The antimicrobial activity was assessed for eight bacterial (Bacillus cereus, Staphylococcus aureus, Salmonella enterica subsp. enterica, Proteus vulgaris, Enterococcus faecalis, Escherichia coli, Pseudomonas aeruginosa and Listeria monocytogenes), five moulds (Aspergillus flavus, Aspergillus niger, Aspergillus ochraceus, Penicillium citrinum, Penicillium expansum) and one yeast strain (Saccharomyces cerevisiae). The agar diffusion method revealed the blackberry GTE as the most effective since it inhibited the growth of three bacterial, four moulds and one yeast species, having considered the total number of affected microorganism species. Next to the blackberry, the olive GTE appeared to be the second most efficient, suppressing five bacterial strains but no moulds or yeasts. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) were then determined for each GTE and the microorganisms tested. Noticeably, the olive GTE appeared to feature the strongest bacteriostatic and bactericidal outcome, displaying specificity for S. aureus, E. faecalis and L. monocytogenes. The other GTEs, such as blueberry, walnut, black mulberry and almond (the list indicates relative strength), were more effective at suppressing microbial growth than inducing microbial death. However, some species specificities were also evident, while the blackcurrant GTE had no significant antimicrobial activity. Having seen the antimicrobial properties of the analysed GTEs, especially the olive and black mulberry GTEs, these could be envisioned as potential antimicrobials that might enhance antibiotic therapies efficiency, while the blackberry GTE would act as an antifungal agent. Some of the GTE mixtures analysed have shown interesting antimicrobial synergies, and all the antimicrobial effects observed argue for extending these studies to include pathological microorganisms. Full article

Altogether, 39 cereal flour samples taken from a Serbian market were analyzed for mycobiota and mycotoxin content, among which were six Triticum aestivum specimens, five Triticum dicoccum specimens, four Hordeum vulgare specimens, five Fagopyrum esculentum specimens, three Secale cereale specimens, five Triticum spelta specimens, four Avena sativa specimens, two Oryza sativa specimens, two Zea mays specimens, and one specimen each of Panicum miliaceum, Triticum monococcum, and Triticum turgidum ssp. turanicum. To determine the mycobiota content using dilution techniques, the flour samples were transferred to a non-selective DG18 nutrient. The number of coloniforming units (CFU/g) varied from less than 100 (in the case of five samples, namely, two O. sativa, and one specimen each of S. cereale, H. vulgare, and T. aestivum) to as high as 5000 CFU/g (S. cereale), 6000 (A. sativa), 11,000 (T. aesticum), and 40,000 (Z. mays). The identification of fungal genera and species was performed on Czapex-Dox Agar and Potato dextrose Agar on the basis of the isolates’ colony characteristics and the morphology of the examined reproductive organs. The isolated fungi belonged to the following genera: Aspergillus, Penicillium, Alternaria, and Fusarium. Species from these genera are well-known mycotoxin-producing fungi. Among the identified species were A. candidus, A. flavus, A. carbonarius, A. ochraceus, A. oryzae, P. solitum, P. citrinum, P. griseofulvum, P. brevicompactum, A. alternata, F. avenaceum, and F. graminearum. The mycotoxin content was determined via the ELISA technique using Eurofins Technologies Hungary KFT kits for aflatoxin B1, deoxynivalenol, total aflatoxins, ochratoxin A, and zearalenone. In the case of eighteen samples, the total aflatoxin content was above the limit of detection, and seven of these samples were contaminated with aflatoxin B1, eight were contaminated with ochratoxin A, two were contaminated with dexynivalenol, and one was contaminated with zearalenon. Two samples of T. aestivum were contaminated with one or more toxins (33%), and the number of samples contaminated three for T. dicoccum (60%), one for H. vulgare (25%), four for F. esculentum (80%), one for S. cereale (33%), two for T. spelta (40%), three for A. sativa (75%), two for O. sativa (100%), two for Z. mays (100%), one for P. miliaceum (100%), one for T. monococcum (100%), and one for T. turgidum ssp. turanicum (100%). Full article

In the context of the mysterious Balkan endemic nephropathy of the 1900s, and the discovery in the 1960s of the potent mycotoxin ochratoxin A, experimental research projects sought to explore any inter-relationship. Experimental lifetime administration of the toxin to male rats had revealed renal DNA adducts with the toxin, correlated with renal tumours, confirmation of which required molecular evidence. Consequently, production of ¹⁴C-ochratoxin A of a high specific radioactivity was required, practical biosynthetic detail of which had not previously been published. A fermentation study of Aspergillus ochraceous was carried out during 2002 for a European project, to select for the production of high-quality ¹⁴C-ochratoxin A, necessarily exploring for the maximum diversion of ¹⁴C-sodium acetate into the pentaketide portion of mycotoxin. Experimentation necessarily had to optimise the competitive context of fungal growth dynamics and addition of the biosynthetic precursor in the early days of shaken-flask fermentation before adding the radiolabelled precursor. From optimal fermentation, 50 mg of the ¹⁴C ochratoxin A was supplied within a European project for DNA adduct experimentation, but that proved negative as subsequently published. Experimental description of the radiolabelled ochratoxin A production was later made in a doctoral thesis, but is first publicised here. Further review of the literature reveals an explanation for the published failure to confirm rat DNA/ochratoxin A adduct formation, for which further experimentation is now recommended. Full article

Biotransformation of ursonic acid (1) by two fungal strains Aspergillus ochraceus CGMCC 3.5324 and Aspergillus oryzae CGMCC 3.407 yielded thirteen new compounds (4, 5, 7–10, and 13–19), along with five recognized ones. The structural details of new compounds were determined through spectroscopic examination (NMR, IR, and HR-MS) and X-ray crystallography. Various modifications, including hydroxylation, epoxidation, lactonization, oxygen introduction, and transmethylation, were identified on the ursane core. Additionally, the anti-neuroinflammatory efficacy of these derivatives was assessed on BV-2 cells affected by lipopolysaccharides. It was observed that certain methoxylated and epoxylated derivatives (10, 16, and 19) showcased enhanced suppressive capabilities, boasting IC₅₀ values of 8.2, 6.9, and 5.3 μM. Such ursonic acid derivatives might emerge as potential primary molecules in addressing neurodegenerative diseases. Full article

Aspergillus parasiticus and Aspergillus ochraceus are important pathogenic fungi that pose a serious threat because of their ability to produce mycotoxins, including ochratoxin A (OTA) and aflatoxins (AFs). The main method of reducing these pathogens is the use of chemical fungicides, though recently there has been a focus on finding biological control agents. The obtained results from this study indicate the great potential of two wild yeast strains, Aureobasidium pullulans PP3 and Saitozyma podzolicus D10, in the biological control of A. parasiticus and A. ochraceus and reductions in the amount of OTA and AFs they produce. In vitro, the growth of the mycelium of pathogens was reduced by 41.21% to 53.64%, and spore germination was inhibited by 58.39% to 71.22%. Both yeast strains produced the enzymes chitinase, β-1,3-glucanase, and amylase, and A. pullulans PP3 additionally produced protease and cellulase. This yeast strain also had the ability to grow over a wide range of temperature (4–30 °C), salinity (0–12%) and pH (4–11) conditions. No growth of the yeast was observed at 37 °C, nor any biogenic amines or hydrogen sulfide production. Adding the tested yeast inoculum to the dough reduced OTA (within 14.55–21.80%) and AFs (within 18.10–25.02%) in the model bread. Full article

Biflavonoids are dimeric forms of flavonoids that have recently gained importance as an effective new scaffold for drug discovery. In particular, 3′-8″-biflavones exhibit antiviral and antimicrobial activity and are promising molecules for the treatment of neurodegenerative and metabolic diseases as well as cancer therapies. In the present study, we directly compared 3′-8″-biflavones (amentoflavone, bilobetin, ginkgetin, isoginkgetin, and sciadopitysin) and their monomeric subunits (apigenin, genkwanin, and acacetin) and evaluated their radical scavenging activity (with DPPH), antifungal activity against mycotoxigenic fungi (Alternaria alternata, Aspergillus flavus, Aspergillus ochraceus, Fusarium graminearum, and Fusarium verticillioides), and inhibitory activity on enzymes (acetylcholinesterase, tyrosinase, α-amylase, and α-glucosidase). All the tested compounds showed weak radical scavenging activity, while antifungal activity strongly depended on the tested concentration and fungal species. Biflavonoids, especially ginkgetin and isoginkgetin, proved to be potent acetylcholinesterase inhibitors, whereas monomeric flavonoids showed higher tyrosinase inhibitory activity than the tested 3′-8″-biflavones. Amentoflavone proved to be a potent α-amylase and α-glucosidase inhibitor, and in general, 3′-8″-biflavones showed a stronger inhibitory potential on these enzymes than their monomeric subunits. Thus, we can conclude that 3′-8″-dimerization enhanced acetylcholinesterase, α-amylase, and α-glucosidase activities, but the activity also depends on the number of hydroxyl and methoxy groups in the structure of the compound. Full article