Ether-Diol Ambiguity: An Inconspicuous Issue in the Structure Elucidation of Oxygenated Natural Products.

Tertiary and allylic hydroxyl groups readily eliminate water during positive ion mode mass spectrometry and may show similar NMR spectra to their corresponding ethers. In a routine structure elucidation workflow, these factors can cause researchers to incorrectly assign diol moieties as ethers or vice versa, leading to inaccurate chemical structures. After facing this problem during our work on oxygenated sesquiterpenoids from a Fusarium sp. fungal strain, we became aware of this challenging issue. We examined the literature for oxygenated natural products bearing these functional groups, and with the aid of density functional calculations of NMR chemical shifts, we now report the structures of 15 natural products that should be revised. We further establish that derivatizing sub-micromolar amounts of alcohols to their sulfates can be used to distinguish these from their corresponding ethers using liquid chromatography negative ion mode mass spectrometry. Finally, we isolated lignoren/cyclonerodiol from the Fusarium sp. culture extract and supported its revised identity as cyclonerodiol using this sulfation approach. Our results suggest that ether-diol ambiguity could be a prevalent issue affecting the structure elucidation of oxygenated natural products and highlight the importance of using complementary techniques, such as sulfation with LC-(-)-ESI-MS or density functional calculations of NMR chemical shifts.

The occurrence and biological control of zearalenone in cereals and cereal-based feedstuffs: a review.

Zearalenone, a prominent mycotoxin produced by Fusarium spp., ubiquitously contaminates cereal grains and animal feedstuffs. The thermal stability of zearalenone creates serious obstacles for traditional removal methods, which may introduce new safety issues, or reducing nutritional quality. In contrast, biological technologies provide appealing benefits such as easy to apply and effective, with low toxicity byproducts. Thus, this review aims to describe the occurrence of zearalenone in cereals and cereal-based feedstuffs in the recent 5 years, outline the rules and regulations regarding zearalenone in the major countries, and discuss the recent developments of biological methods for controlling zearalenone in cereals and cereal-based feedstuffs. In addition, this article also reviews the application and the development trend of biological strategies for removal zearalenone in cereals and cereal-based feedstuffs.

Mycotoxin Accumulation in Dry Rot Potato Tubers from Algeria and Toxigenic Potential of Associated Isolates of Fusarium Genus.

The presence of different mycotoxins in 232 tuber samples exhibiting dry rot symptoms and their associated Fusarium strains from two production sites in Algeria was investigated. LC-MS/MS was used to simultaneously detect and quantify 14 mycotoxins, including trichothecenes and non-trichothecenes. A total of 49 tubers were contaminated with at least one mycotoxins, including T-2, HT-2, Diacetoxyscirpenol (DAS), 15-acetoxyscirpenol (15-AS) and Beauvericin (BEA). Positive samples from the Bouira region had a significantly higher level of toxin contamination compared to Ain Defla (56.34% and 5.59%, respectively). A total of 283 Fusarium strains were isolated: 155 from Bouira and 128 from Ain Defla. These strains were evaluated for their ability to produce the targeted mycotoxins. The results showed that 61.29% and 53.9% of strains originate from Bouira and Ain Defla regions were able to produce Nivalenol, Fusarenone-X, DAS, 15-AS, Neosolaniol, BEA and Zearalenone. The phylogenetic analysis of the conserved ribosomal internal transcribed spacer (ITS) sequences of 29 Fusarium strains, representative of the recorded mycotoxins profiles, was distributed into 5 Fusarium species complexes (SC): F. incarnatum-equiseti SC (FIESC), F. sambucinum SC (FSAMSC), F. oxysporum SC (FOSC), F. tricinctum SC (FTSC) and F. redolens SC (FRSC). This is the first study determining multiple occurrences of mycotoxins contamination associated to Fusarium dry rot of potato in Algeria and highlights fungal potential for producing trichothecene and non-trichothecens mycotoxins.

Co-occurrence of mycotoxins in stored maize from southern and southwestern Ethiopia.

Maize grain samples collected from 129 small-scale farmers' stores in southern and southwestern Ethiopia were analysed by LC-MS/MS for a total of 218 mycotoxins and other fungal metabolites of which 15% were regulated mycotoxins. Mycotoxins produced by Penicillium, Aspergillus, and Fusarium accounted for 31%, 17%, and 12% of the metabolites, respectively. Most of the current samples were contaminated by masked and/or emerging mycotoxins with moniliformin being the most prevalent one, contaminating 93% of the samples. Each sample was co-contaminated by 3 to 114 mycotoxins/fungal metabolites. Zearalenone, fumonisin B1, and deoxynivalenol were the dominant mycotoxins, occurring in 78%, 61%, and 55% of the samples with mean concentrations of 243, 429, and 530 µg/kg, respectively. The widespread co-occurrence of several mycotoxins in the samples may pose serious health risks due to synergistic/additional effects.

Surface-Imprinted Polysiloxane with Recognition Ability Based on an ITO Layer for Rapid Detection of Fusarium oxysporum f. sp. cubense by the Naked Eye.

Direct observation by the naked eye of fluorescence-stained microbes adsorbed on surface imprinted polymers (SIPs) is highly challenging and limited by speed, accuracy and the semiquantitative nature of the method. In this study, we tested for the presence of spores of Fusarium oxysporum f. sp. cubense race 4 (Foc4), which cause severe banana Fusarium wilt disease and reduces the area of banana plants. This kind of spore can become dormant in soil, which means that the detection of secreted molecules (molecular imprinting) in soil may be inaccurate; detection methods such as polymerase chain reaction (PCR) and Raman spectroscopy are more accurate but time-consuming and inconvenient. Therefore, a semiquantitative and rapid SIP detection method for Foc4 was proposed. Based on the ITO conductive layer, a reusable and naked-eye-detectable Foc4-PDMS SIP film was prepared with a site density of approximately 9000 mm-2. Adsorption experiments showed that when the Foc4 spore concentration was between 104 to 107 CFU/mL, the number of Foc4 spores adsorbed and the fluorescence intensity were strongly correlated with the concentration and could be fully distinguished by the naked eye after fluorescence staining. Adsorption tests on other microbes showed that the SIP film completely recognized only the Foc series. All the results were highly consistent with the naked-eye observations after fluorescence staining, and the results of the Foc4-infected soil experiment were also close to the ideal situation. Taken together, these results showed that Foc4-PDMS SIPs have the ability to rapidly and semiquantitatively detect the concentration of Foc in soil, which can provide good support for banana cultivation. This method also has potential applications in the detection of other fungal diseases.

A novel strategy for detoxification of deoxynivalenol via modification of both toxic groups.

Deoxynivalenol (DON) is the most abundant mycotoxin in cereal crops and derived foods and is of great concern in agriculture. Bioremediation strategies have long been sought to minimize the impact of mycotoxin contamination, but few direct and effective enzyme-catalyzed detoxification methods are currently available. In this study, we established a multi-enzymatic cascade reaction and successfully achieved detoxification at double sites: glutathionylation for the C-12,13 epoxide group and epimerization for the C-3 hydroxyl group. This yielded novel derivatives of DON, 3-epi-DON-13-glutathione (3-epi-DON-13-GSH) as well as its by-product, 3-keto-DON-13-GSH, for which precise structures were validated via liquid chromatography-high-resolution tandem mass spectrometry (LC-HRMS) and nuclear magnetic resonance (NMR) spectroscopy. Both cell viability and DNA synthesis assays demonstrated dramatically decreased cytotoxicity of the double-site modified product 3-epi-DON-13-GSH. These findings provide a promising and urgently needed novel method for addressing the problem of DON contamination in agricultural and industrial settings.

Secofumitremorgins C and D, a pair of atropisomers from saltern-derived fungus Aspergillus fumigatus GXIMD00544.

A pair of atropisomers secofumitremorgins C (1a) and D (1b), together with fifteen known alkaloids (2-16), were isolated from a saltern-derived fungus Aspergillus fumigatus GXIMD00544. The structures of atropisomers 1a and 1b were elucidated by the detailed spectroscopic data, chemical reaction and quantum chemical calculations. Compounds 1 and 8 displayed antifungal spore germination effects against plant pathogenic fungus associated with sugarcane Fusarium sp. with inhibitory rates of 53% and 77% at the concentration of 100 µM, repectively. Atropisomers 1 also exhibited antifouling potential against Balanus amphitrite larval settlement with an inhibitory rate of 96% at the concentration of 100 µM.

Mycoproteins and their health-promoting properties: Fusarium species and beyond.

Filamentous fungal mycoproteins have gained increasing attention as sustainable alternatives to animal and plant-based proteins. This comprehensive review summarizes the nutritional characteristics, toxicological aspects, and health-promoting effects of mycoproteins, focusing on those derived from filamentous fungi, notably Fusarium venenatum. Mycoproteins are characterized by their high protein content, and they have a superior essential amino acid profile compared to soybeans indicating excellent protein quality and benefits for human nutrition. Additionally, mycoproteins offer enhanced digestibility, further highlighting their suitability as a protein source. Furthermore, mycoproteins are rich in dietary fibers, which have been associated with health benefits, including protection against metabolic diseases. Moreover, their fatty acids profile, with significant proportions of polyunsaturated fatty acids and absence of cholesterol, distinguishes them from animal-derived proteins. In conclusion, the future of mycoproteins as a health-promoting protein alternative and the development of functional foods relies on several key aspects. These include improving the acceptance of mycoproteins, conducting further research into their mechanisms of action, addressing consumer preferences and perceptions, and ensuring safety and regulatory compliance. To fully unlock the potential of mycoproteins and meet the evolving needs of a health-conscious society, continuous interdisciplinary research, collaboration among stakeholders, and proactive engagement with consumers will be vital.

Mycotoxin contamination in organic and conventional cereal grain and products: A systematic literature review and meta-analysis.

There is still considerable controversy about the relative risk of mycotoxin exposure associated with the consumption of organic and conventional cereals. Using validated protocols, we carried out a systematic literature review and meta-analyses of data on the incidence and concentrations of mycotoxins produced by Fusarium, Claviceps, Penicillium, and Aspergillus species in organic and conventional cereal grains/products. The standard weighted meta-analysis of concentration data detected a significant effect of production system (organic vs. conventional) only for the Fusarium mycotoxins deoxynivalenol, with concentrations ∼50% higher in conventional than organic cereal grains/products (p < 0.0001). Weighted meta-analyses of incidence data and unweighted meta-analyses of concentration data also detected small, but significant effects of production system on the incidence and/or concentrations of T-2/HT-2 toxins, zearalenone, enniatin, beauvericin, ochratoxin A (OTA), and aflatoxins. Multilevel meta-analyses identified climatic conditions, cereal species, study type, and analytical methods used as important confounding factors for the effects of production system. Overall, results from this study suggest that (i) Fusarium mycotoxin contamination decreased between the 1990s and 2020, (ii) contamination levels are similar in organic and conventional cereals used for human consumption, and (iii) maintaining OTA concentrations below the maximum contamination levels (3.0 µg/kg) set by the EU remains a major challenge.

[Active secondary metabolites from an endophytic fungus Fusarium solani MBM-5 of Datura arborea].

This study investigated the chemical and biological activity of the secondary metabolites from an endophytic fungus Fusa-rium solani MBM-5 of Datura arborea. A total of six alkenoic acid compounds, including a new compound and five known ones, were isolated from the ethyl acetate extract of F. solani MBM-5 by using the chromatographic methods(open ODS column chromatography, silica gel column chromatography, Sephadex LH-20, and semi-preparative HPLC). The structures of the compounds were identified by using their physical and chemical data, spectroscopic methods(UV, IR, NMR, and HR-ESI-MS), and Mosher's reaction, which were fusaridioic acid E(1), fusaridioic acid C(2), fusaridioic acid A(3), L660282(4), hymeglusin(5), and hymeglnone(6). Compound 1 is new. MTT assay and Griss method were used to evaluate the growth inhibition of all the compounds against two tumor cells, as well as their influence and anti-inflammatory action on the release of NO from LPS-induced RAW264.7 cells. The results showed that compound 5 had strong growth inhibition activity against A549 and HepG2 cell lines, with IC_(50) values of 4.70 and 13.57 µmol·L~(-1), respectively. Compounds 1 and 6 significantly inhibited the release of NO from LPS-induced RAW264.7 cells, with IC_(50) values of 77.00 and 70.33 µmol·L~(-1), respectively.

The study of iron- and copper-binding proteome of Fusarium oxysporum and its effector candidates.

Fusarium oxysporum f.sp. lycopersici is a phytopathogen which causes vascular wilt disease in tomato plants. The survival tactics of both pathogens and hosts depend on intricate interactions between host plants and pathogenic microbes. Iron-binding proteins (IBPs) and copper-binding proteins (CBPs) play a crucial role in these interactions by participating in enzyme reactions, virulence, metabolism, and transport processes. We employed high-throughput computational tools at the sequence and structural levels to investigate the IBPs and CBPs of F. oxysporum. A total of 124 IBPs and 37 CBPs were identified in the proteome of Fusarium. The ranking of amino acids based on their affinity for binding with iron is Glu > His> Asp > Asn > Cys, and for copper is His > Asp > Cys respectively. The functional annotation, determination of subcellular localization, and Gene Ontology analysis of these putative IBPs and CBPs have unveiled their potential involvement in a diverse array of cellular and biological processes. Three iron-binding glycosyl hydrolase family proteins, along with four CBPs with carbohydrate-binding domains, have been identified as potential effector candidates. These proteins are distinct from the host Solanum lycopersicum proteome. Moreover, they are known to be located extracellularly and function as enzymes that degrade the host cell wall during pathogen-host interactions. The insights gained from this report on the role of metal ions in plant-pathogen interactions can help develop a better understanding of their fundamental biology and control vascular wilt disease in tomato plants.

Proliferacorins A-M: Thirteen undescribed acorane sesquiterpenoids isolated from the fungus Fusarium proliferatum.

In this study, 13 previously undescribed acorane sesquiterpenoids, namely, proliferacorins A-M, were isolated from the solid fermentation of Fusarium proliferatum. Their structures and absolute configurations were confirmed via spectroscopic analyses, quantum-chemical NMR calculations with DP4+ probability analyses, ECD calculations and comparisons, and single-crystal X-ray diffraction techniques. Proliferacorins A-E (1-5) have a 7-oxa-tricyclo[6.3.1.01,5]tridecane decorated with a rare ether bridge between C-7 and C-11, while proliferacorin F (6) possesses a 7-oxa-tricyclic[6.4.0.01,5]dodecane skeleton with an unusual ether bond between C-6 and C-11. Proliferacorins C and D (3 and 4) are a pair of isomers at the carbon bridge between C-5 and C-7, whereas proliferacorins H and I (8 and 9) are a pair of spiro carbon isomers. All isolates were tested for their cytotoxic, anti-inflammatory, and immunosuppressive activities.

Effect of Fungicide Application and Corn Hybrid Class on the Presence of Fusarium graminearum and the Concentration of Deoxynivalenol in Ear and Stalk Parts of Corn (Zea mays) Used for Silage.

In Wisconsin, the use of brown midrib (BMR) corn (Zea mays) hybrids for ensiling and subsequent feeding to dairy cows is quite common. The overall milk production from cows fed silage from BMR hybrids is typically higher than those fed silage made from dual-purpose hybrids. Gibberella diseases (ear and stalk rot) caused by Gibberella zeae (anamorph; Fusarium graminearum) and the accompanying accumulation of the mycotoxin deoxynivalenol (DON) can be significant issues during the field production of BMR hybrids. The work presented here aimed to understand the role of hybrid class on the distribution of F. graminearum DNA and DON in the ear and stalk parts of corn for silage. An ear and stalk partitioned sample experiment was conducted on silage corn from field trials in Arlington, Wisconsin, in 2020 and 2021. The trials were arranged in a randomized complete block design in both years, including one BMR hybrid, one dual-purpose hybrid, and seven fungicide application regimes. Paired ear and stalk samples were physically separated, dried, and ground at harvest before determining the concentration of F. graminearum DNA and DON in each sample. Across both years, the main effects of hybrid, treatment, and plant part were not significant (P > 0.1) on DON concentration. However, the hybrid-by-plant part interaction effect was significant (P < 0.01). Ears of the BMR hybrid accumulated the most DON, whereas the dual-purpose hybrid ears had the lowest DON concentration. The concentrations of DON and F. graminearum DNA were significantly (P < 0.01) and highly correlated in the ear (r = 0.73) but not in the stalk (r = 0.09, P = 0.33). These findings suggest that DON accumulation in the corn ear is a major contributor in the difference observed in the total DON between the hybrid classes. Therefore, growers and researchers are encouraged to focus production and breeding on hybrids in both classes that accumulate less DON in ears, resulting in lower total DON in corn chopped for silage.

Chemical Constituents of a Marine-derived Fungus Fusarium oxysporum F0888 and their Antibacterial Activity.

Five new compounds, including four hydroxyphenylacetic acid derivatives, stachylines H-K (1-4), a derivative of hydroxyphenylethanol (5), as well as seven known compounds were obtained from a marine-derived fungus Fusarium oxysporum F0888 isolated from sediments in the South China Sea. The structures and absolute configurations of new compounds were determined by spectroscopic (IR, NMR, and HR-ESI-MS) analyses, comparison of optical rotations, and the modified Mosher's MTPA ester method. Antimicrobial and anti-inflammatory activities of compounds 1-12 were tested. Unfortunately, all of isolated compounds were inactivity.

A novel core-shell up-conversion nanoparticles immunochromatographic assay for the detection of deoxynivalenol in cereals.

Deoxynivalenol (DON) toxin is a type B group of trichothecene mycotoxins mainly originating from specific Fusarium fungi, seriously harming human and livestock health. Herein, a novel core-shell up-conversion nanoparticles immunochromatographic assay (CS-UCNPs-ICA) was developed for deoxynivalenol based on the competitive reaction principle. By exploiting the fluorescence intensity of the T and C lines of CS-UCNPs-ICA, the concentrations of DON were obtained sensitively and precisely under optimized conditions in 5 min with a detection limit of 0.1 ng/mL. The CS-UCNPs-ICA strips only specifically detect DON and its derivatives (3-Ac-DON and 15-Ac-DON), with no cross-reaction with other mycotoxins. The low CV values illustrated a modest intra- and inter-assay variation, confirming the superior precision of this method. In the spiked experiment, the mean recoveries of corn and wheat ranged from 94.74% to 100.90% and 96.21%-104.81%, respectively. Furthermore, the approach generated results that were in good agreement with data from HPLC and ELISA analyses of naturally contaminated feed and cereals, confirming that the significant advantages of proposed strips were their high practicality, rapidness, and simplicity. Therefore, the CS-UCNPs-ICA strips platform serves as a promising candidate for developing new approaches for rapid testing or high throughput screening from DON in food products.

Improving the productivity of gibberellic acid by combining small-molecule compounds-based targeting technology and transcriptomics analysis in Fusarium fujikuroi.

Gibberellic acid (GA3), produced industrially by Fusarium fujikuroi, stands as a crucial plant growth regulator extensively employed in the agriculture filed while limited understanding of the global metabolic network hinders researchers from conducting rapid targeted modifications. In this study, a small-molecule compounds-based targeting technology was developed to increase GA3 production. Firstly, various small molecules were used to target key nodes of different pathways and the result displayed that supplement of terbinafine improved significantly GA3 accumulation, which reached to 1.08 g/L. Subsequently, lipid and squalene biosynthesis pathway were identified as the key pathways influencing GA3 biosynthesis by transcriptomic analysis. Thus, the strategies including in vivo metabolic engineering modification and in vitro supplementation of lipid substrates were adopted, both contributed to an enhanced GA3 yield. Finally, the engineered strain demonstrated the ability to achieve a GA3 yield of 3.24 g/L in 5 L bioreactor when utilizing WCO as carbon source and feed.

Toxicity of the emerging mycotoxins beauvericin and enniatins: A mini-review.

Beauvericin and enniatins, emerging mycotoxins produced mainly by Fusarium species, are natural contaminants of cereals and cereal products. These mycotoxins are cyclic hexadepsipeptides with ionophore properties and their toxicity mechanism is related to their ability to transport cations across the cell membrane. Beauvericin and enniatins are cytotoxic, as they decrease cell viability, promote cell cycle arrest, and increase apoptosis and the generation of reactive oxygen species in several cell lines. They also cause changes at the transcriptomic level and have immunomodulatory effects in vitro and in vivo. Toxicokinetic results are scarce, and, despite its proven toxic effects in vitro, no regulation or risk assessment has yet been performed due to a lack of in vivo data. This mini-review aims to report the information available in the literature on studies of in vitro and in vivo toxic effects with beauvericin and enniatins, which are mycotoxins of increasing interest to animal and human health.

The antifungal metabolites isolated from maize endophytic fungus Fusarium sp. induced by OSMAC strategy.

Six new sesquiterpenes, fusarchlamols A-F (1, 2, 4-7); one new natural product of sesquiterpenoid, methyltricinonoate (3); and ten known compounds were found from Fusarium sp. cultured in two different media by the one strain many compounds strategy. The compounds (1, 2, and 4-11) were isolated from Fusarium sp. in PDB medium, and compounds (3-5, 8, and 10-17) were discovered from Fusarium sp. in coffee medium. Additionally, the configuration of 8 was first reported in the research by Mosher's method. The structures were established by 1D, 2D NMR, mass spectrometry, calculated ECD spectra, and Mosher's method. Compounds 1, 2, 6/7, 12, and 16 indicated significant antifungal activities against the phytopathogen Alternaria alternata isolated from Coffea arabica with MICs of 1 µg/mL. The investigation on the anti-phytopathogen activity of metabolites can provide lead compounds for agrochemicals.

Mid-Level Data Fusion Combined with the Fingerprint Region for Classification DON Levels Defect of Fusarium Head Blight Wheat.

In this study, a method of mid-level data fusion with the fingerprint region was proposed, which was combined with the characteristic wavelengths that contain fingerprint information in NIR and FT-MIR spectra to detect the DON level in FHB wheat during wheat processing. NIR and FT-MIR raw spectroscopy data on normal wheat and FHB wheat were obtained in the experiment. MSC was used for pretreatment, and characteristic wavelengths were extracted by CARS, MGS and XLW. The variables that can effectively reflect fingerprint information were retained to build the mid-level data fusion matrix. LS-SVM and PLS-DA were applied to investigate the performance of the single spectroscopic model, mid-level data fusion model and mid-level data fusion with fingerprint information model, respectively. The experimental results show that mid-level data fusion with a fingerprint information strategy based on fused NIR and FT-MIR spectra represents an effective method for the classification of DON levels in FHB wheat samples.

Occurrence of resorcyclic acid lactones in porcine urine: discrimination between illegal use and contamination.

Zeranol (α-zearalanol, α-ZAL), is a resorcyclic acid lactone (RAL). Its administration to farm animals to improve meat production has been prohibited in the European Union due to the potential risk to human health. However, it has been demonstrated that α-ZAL may be present in livestock animals due to Fusarium fungi that produce fusarium acid lactones contamination in feed. The fungi produce a small amount of zearalenone (ZEN), which is metabolized to zeranol. The potential endogenous origin of α-ZAL makes it difficult to correlate positive samples to a potential illicit treatment with α-ZAL. We present two experimental studies that investigated the origin of natural and synthetic RALs in porcine urine. Urine samples from pigs that were either fed with ZEN-contaminated feed or administered α-ZAL by injection were analyzed by liquid chromatography coupled to tandem mass spectrometry, with the method validated according to Commission Implementing Regulation (EU) 2021/808. The data show that although the concentration of α-ZAL in the ZEN feed-contaminated samples is significantly lower than in the illicit administration samples, α-ZAL can occur in porcine urine via natural metabolism. Additionally, the feasibility of using the ratio of forbidden/fusarium RALs in porcine urine as a reliable biomarker for illicit treatment with α-ZAL administration was evaluated for the first time. This study demonstrated that the obtained ratio in the contaminated ZEN feed study was close to 1, while in the illegally administered α-ZAL samples the ratio is always higher than 1 (up to 135). Therefore, this study proves that the ratio criteria (already used when a forbidden RAL is detected in bovine urine) may also be used for porcine urine.