Fusarium graminearum Species Complex: A Bibliographic Analysis and Web-Accessible Database for Global Mapping of Species and Trichothecene Toxin Chemotypes.

Fusarium graminearum is ranked among the five most destructive fungal pathogens that affect agroecosystems. It causes floral diseases in small grain cereals including wheat, barley, and oats, as well as maize and rice. We conducted a systematic review of peer-reviewed studies reporting species within the F. graminearum species complex (FGSC) and created two main data tables. The first contained summarized data from the articles including bibliographic, geographic, methodological (ID methods), host of origin and species, while the second data table contains information about the described strains such as publication, isolate code(s), host/substrate, year of isolation, geographical coordinates, species and trichothecene genotype. Analyses of the bibliographic data obtained from 123 publications from 2000 to 2021 by 498 unique authors and published in 40 journals are summarized. We describe the frequency of species and chemotypes for 16,274 strains for which geographical information was available, either provided as raw data or extracted from the publications, and sampled across six continents and 32 countries. The database and interactive interface are publicly available, allowing for searches, summarization, and mapping of strains according to several criteria including article, country, host, species and trichothecene genotype. The database will be updated as new articles are published and should be useful for guiding future surveys and exploring factors associated with species distribution such as climate and land use. Authors are encouraged to submit data at the strain level to the database, which is accessible at https://fgsc.netlify.app.

Genetic structure of Fusarium verticillioides populations from maize in Iran.

Fusarium verticillioides is one of the most important fungal pathogens of maize since it causes severe yield losses and produces the mycotoxins fumonisins that represent a major concern for human and animal health. Information about genetic diversity and population structure of fungal pathogens is essential for developing disease management strategies. The aim of this research was to investigate the genetic structure of F. verticillioides isolated from different provinces of Iran through determination of mating type idiomorphs, phylogenetic analyses based on translation elongation factor-1 alpha (EF-1α), RNA Polymerase II Subunit (RPB2), beta-tubulin (tub2) and Calmodulin (cmdA) genes and genetic diversity analyses based on 6 simple-sequence repeats (SSRs). Both mating types were detected in Iranian populations of F. verticillioides, particularly in Qazvin and Khuzestan, with equal frequency, which highlighted that sexual reproduction is favorable under field conditions. However, the linkage disequilibrium indices did not support the hypothesis of random mating in Khuzestan and Fars. Although assessment of nucleotide diversity based on housekeeping genes showed low level of variation among strains, genotype diversity based on SSRs revealed a high level of genetic diversity within Iranian populations. AMOVA analysis highlighted that the genetic variation of F. verticillioides in Iran was mainly distributed within population of a single area (97%), while a small proportion of genetic variation (3%) resided among populations. These patterns of variation are likely explained by the continuous gene flow among populations isolated from different areas. On the other hand, principal coordinate analysis indicated that the distribution of genetic variation among populations could be explained by the geographical distances. Consequently, to reduce pathogen gene flow among regions, the quarantine processes in Iran should be intensified.

Antioxidant and Pro-Oxidant Capacities as Mechanisms of Photoprotection of Olive Polyphenols on UVA-Damaged Human Keratinocytes.

A wide variety of polyphenols are reported to have considerable antioxidant and skin photoprotective effects, although the mechanisms of action are not fully known. Environmentally friendly and inexpensive sources of natural bioactive compounds, such as olive mill wastewater (OMWW), the by-product of olive-oil processing, can be considered an economic source of bioactive polyphenols, with a range of biological activities, useful as chemotherapeutic or cosmeceutical agents. Green strategies, such as the process based on membrane technologies, allow to recover active polyphenols from this complex matrix. This study aims to evaluate the antioxidant, pro-oxidant, and photoprotective effects, including the underlying action mechanism(s), of the ultra-filtered (UF) OMWW fractions, in order to substantiate their use as natural cosmeceutical ingredient. Six chemically characterized UF-OMWW fractions, from Italian and Greek olive cultivar processing, were investigated for their antioxidant activities, measured by Trolox Equivalent Antioxidant Capacity (TEAC), LDL oxidation inhibition, and ROS-quenching ability in UVA-irradiated HEKa (Human Epidermal Keratinocytes adult) cultures. The photoprotective properties of UF-OMWW were assayed as a pro-oxidant-mediated pro-apoptotic effect on the UVA-damaged HEKa cells, which can be potentially involved in the carcinogenesis process. All the UF-OMWW fractions exerted an effective antioxidant activity in vitro and in cells when administered together with UV-radiation on HEKa. A pro-oxidative and pro-apoptotic effect on the UVA-damaged HEKa cells were observed, suggesting some protective actions of polyphenol fraction on keratinocyte cell cultures.

The life and times of yeasts in traditional food fermentations.

Yeasts are eukaryotic microorganisms which have a long history in the biotechnology of food production, as they have been used since centuries in bread-making or in the production of alcoholic beverages such as wines or beers. Relative to this importance, a lot of research has been devoted to the study of yeasts involved in making these important products. The role of yeasts in other fermentations in association with other microorganisms - mainly lactic acid bacteria - has been relatively less studied, and often it is not clear if yeasts occurring in such fermentations are contaminants with no role in the fermentation, spoilage microorganisms or whether they actually serve a technological or functional purpose. Some knowledge is available for yeasts used as starter cultures in fermented raw sausages or in the production of acid curd cheeses. This review aimed to summarize the current knowledge on the taxonomy, the presence and potential functional or technological roles of yeasts in traditional fermented plant, dairy, fish and meat fermentations.

FvatfA regulates growth, stress tolerance as well as mycotoxin and pigment productions in Fusarium verticillioides.

FvatfA from the maize pathogen Fusarium verticillioides putatively encodes the Aspergillus nidulans AtfA and Schizasaccharomyces pombe Atf1 orthologous bZIP-type transcription factor, FvAtfA. In this study, a ΔFvatfA deletion mutant was constructed and then genetically complemented with the fully functional FvatfA gene. Comparing phenotypic features of the wild-type parental, the deletion mutant and the restored strains shed light on the versatile regulatory functions played by FvAtfA in (i) the maintenance of vegetative growth on Czapek-Dox and Potato Dextrose agars and invasive growth on unwounded tomato fruits, (ii) the preservation of conidiospore yield and size, (iii) the orchestration of oxidative (H2O2, menadione sodium bisulphite) and cell wall integrity (Congo Red) stress defences and (iv) the regulation of mycotoxin (fumonisins) and pigment (bikaverin, carotenoid) productions. Expression of selected biosynthetic genes both in the fumonisin (fum1, fum8) and the carotenoid (carRA, carB) pathways were down-regulated in the ΔFvatfA strain resulting in defected fumonisin production and considerably decreased carotenoid yields. The expression of bik1, encoding the polyketide synthase needed in bikaverin biosynthesis, was not up-regulated by the deletion of FvatfA meanwhile the ΔFvatfA strain produced approximately ten times more bikaverin than the wild-type or the genetically complemented strains. The abolishment of fumonisin production of the ΔFvatfA strain may lead to the development of new-type, biology-based mycotoxin control strategies. The novel information gained on the regulation of pigment production by this fungus can be interesting for experts working on new, Fusarium-based biomass and pigment production technologies. Key points • FvatfA regulates vegetative and invasive growths of F. verticillioides. • FvatfA also orchestrates oxidative and cell wall integrity stress defenses. • The ΔFvatfA mutant was deficient in fumonisin production. • FvatfA deletion resulted in decreased carotenoid and increased bikaverin yields.

A loop-mediated isothermal amplification (LAMP) assay for rapid detection of fumonisin producing Aspergillus species.

Fumonisins contamination of food commodities is a worldwide problem, especially for maize. The ability to produce fumonisinsis a trait of several species of Fusarium, mainly F. verticillioides and F. proliferatum on maize, and some Aspergillus species. A. niger and its sister species A. welwitschiae, can contribute to fumonisin B2 (FB2) accumulation in maize kernels, although to a lesser extent than fumonisin-producing Fusarium species. Fumonisins risk monitoring represents an effective strategy in the integrated approach for mycotoxin risk management and reduction. The availability of a user-friendlymolecular assay for the detection oftoxigenic fungal species represents a valuable tool in understanding and managing upcoming mycotoxin contamination. In this study, we developed a LAMP assay, based on the detection of fum10, for a rapid and specific molecular detection of FB2-producing A. niger and A. welwistchiae, potentially useful to perform monitoring directly "on site" in maize chain. Results showed that very low amounts of conidia are suitable to detect the presence of the target gene, thus providing information about the presence of FB2-producing Aspergillus species and the possible upcoming fumonisins contamination in maize. The assay was combined with a suitable protocol for "in field" crude DNA extraction and a colorimetric method for easy naked-eye evaluationof results, offering a reliable and user-friendly tool to support effective reduction strategies of mycotoxin contamination in crop management programs.

FvmnSOD is involved in oxidative stress defence, mitochondrial stability and apoptosis prevention in Fusarium verticillioides.

Superoxide dismutases are key enzymes in elimination of the superoxide anion radical (O2 •- ) generated intracellularly or by exogenous oxidative stress eliciting agents, like menadione. In this study, we investigated the physiological role of the manganese superoxide dismutase-encoding gene in Fusarium verticillioides via the construction of a gene deletion mutant, ΔFvmnSOD and comparing its phenotype with that of the wild-type parental strain and a ΔFvmnSOD' C strain, complemented with the functional manganese superoxide dismutase gene. Deletion of FvmnSOD had no effect on the relative intracellular superoxide ratio but increased the sensitivity of the fungus to menadione sodium bisulphite on Czapek-Dox stress agar plates. The lack of FvmnSOD caused changes in mitochondrial morphology and physiology: The volumetric ratio of these cell organelles in the second hyphal segment, as well as the total, the KCN-sensitive cytochrome c-dependent and the KCN+SHAM (salicylhidroxamic acid)-resistant residual respiration rates, were higher in the mutant as compared to the wild-type and the complemented strains. Nevertheless, changes in the respiration rates were attributable to the higher volumetric ratio of mitochondria found in the gene deletion mutant. Changes in the mitochondrial functions also brought about higher sensitivity to apoptotic cell death elicited by the Penicillium chrysogenum antifungal protein. The gene deletion mutant developed significantly thinner hyphae in comparison to the wild-type strain. Deletion of FvmnSOD had no effect on fumonisin B1 and B2 production of the fungus grown in Myro medium as a static culture.

Unravelling the Antifungal Effect of Red Thyme Oil (Thymus vulgaris L.) Compounds in Vapor Phase.

The aim of this work was to evaluate the antifungal activity in vapor phase of thymol, p-cymene, and γ-terpinene, the red thyme essential oil compounds (RTOCs). The Minimum Inhibitory Concentration (MIC) of RTOCs was determined against postharvest spoilage fungi of the genera Botrytis, Penicillium, Alternaria, and Monilinia, by measuring the reduction of the fungal biomass after exposure for 72 h at 25 °C. Thymol showed the lowest MIC (7.0 µg/L), followed by γ-terpinene (28.4 µg/L) and p-cymene (40.0 µg/L). In the case of P. digitatum ITEM 9569, resistant to commercial RTO, a better evaluation of interactions among RTOCs was performed using the checkerboard assay and the calculation of the Fractional Inhibitory Concentration Index (FICI). During incubation, changes in the RTOCs concentration were measured by GC-MS analysis. A synergistic effect between thymol (0.013 ± 0.003 L/L) and γ-terpinene (0.990 ± 0.030 L/L) (FICI = 0.50) in binary combinations, and between p-cymene (0.700 ± 0.010 L/L) and γ-terpinene (0.290 ± 0.010 L/L) in presence of thymol (0.008 ± 0.001 L/L) (FICI = 0.19), in ternary combinations was found. The synergistic effect against the strain P. digitatum ITEM 9569 suggests that different combinations among RTOCs could be defined to control fungal strains causing different food spoilage phenomena.

Characterization of Micronutrients, Bioaccessibility and Antioxidant Activity of Prickly Pear Cladodes as Functional Ingredient.

The Opuntia ficus indica (L.) (OFI) is used as a nutritional and pharmaceutical agent in various dietary and value added products. This study underlines the possible use of native prickly pear cladode powder as a functional ingredient for health-promoting food production. To summarise, chemical characterization of polyphenols, minerals and soluble dietary fibre was performed; furthermore, the antioxidant activity and bioaccessibility of polyphenols and minerals were assessed. Eleven compounds between phenolic acids and flavonoids were identified, with piscidic acid and isorhamnetin derivatives being the most abundant. Opuntia's dietary fibre was mainly constituted of mucilage and pectin, and was composed of arabinose, galactose, glucose, mannose, rhamnose, and xylose sugars. The polyphenols' bioaccessibility was very high: piscidic acid at 200%, eucomic and ferulic acids >110% and flavonoids from 89% to 100%. The prickly pear cladode powder is also a source of minerals, as cations (calcium, sodium, potassium and magnesium) and anions (sulphate and chloride), with high magnesium bioaccessibilty (93%). OFI powder showed good capacity of radical scavenging measured by DPPH and ABTS methods, with 740 and 775 µmol Trolox/100 g OFI, respectively. Finally, the presented results allow the consideration of this natural product as a source of several essential nutrients, with a possible use in the food industry as a functional ingredient.

Microbial quality and safety of milk and milk products in the 21st century.

Milk and milk products have been utilized by humans for many thousands of years. With the advent of metagenomic studies, our knowledge on the microbiota of milk and milk products, especially as affected by the environment, production, and storage parameters, has increased. Milk quality depends on chemical parameters (fat and protein content and absence of inhibitory substances), as well as microbial and somatic cells counts, and affects the price of milk. The effects of hygiene and effective cooling on the spoilage microbiota have shown that proteolytic and lipolytic bacteria such as Pseudomonas or Acinetobacter spp. predominate the spoilage bacterial populations. These bacteria can produce heat-stable proteases and lipases, which remain active after pasteurization and thus can spoil the milk during prolonged storage. Additionally, milk can become contaminated after pasteurization and therefore there is still a high demand on developing better cleaning and sanitation regimes and equipment, as well as test systems to (quantitatively) detect relevant pathogenic or spoilage microorganisms. Raw milk and raw milk cheese consumption is also increasing worldwide with the growing demand of minimally processed, sustainable, healthy, and local foods. In this context, emerging and re-emerging pathogens once again represent a major food safety challenge. As a result of global warming, it is conceivable that not only microbiological risks but also chemical risks relating to presence of mycotoxins or plant toxins in milk will increase. Herein, we provide an overview of the major microbial hazards occurring in the 21st century.

Variation in secondary metabolite production potential in the Fusarium incarnatum-equiseti species complex revealed by comparative analysis of 13 genomes.

BACKGROUND: The Fusarium incarnatum-equiseti species complex (FIESC) comprises 33 phylogenetically distinct species that have been recovered from diverse biological sources, but have been most often isolated from agricultural plants and soils. Collectively, members of FIESC can produce diverse mycotoxins. However, because the species diversity of FIESC has been recognized only recently, the potential of species to cause mycotoxin contamination of crop plants is unclear. In this study, therefore, we used comparative genomics to investigate the distribution of and variation in genes and gene clusters responsible for the synthesis of mycotoxins and other secondary metabolites (SMs) in FIESC. RESULTS: We examined genomes of 13 members of FIESC that were selected based primarily on their phylogenetic diversity and/or occurrence on crops. The presence and absence of SM biosynthetic gene clusters varied markedly among the genomes. For example, the trichothecene mycotoxin as well as the carotenoid and fusarubin pigment clusters were present in all genomes examined, whereas the enniatin, fusarin, and zearalenone mycotoxin clusters were present in only some genomes. Some clusters exhibited discontinuous patterns of distribution in that their presence and absence was not correlated with the phylogenetic relationships of species. We also found evidence that cluster loss and horizontal gene transfer have contributed to such distribution patterns. For example, a combination of multiple phylogenetic analyses suggest that five NRPS and seven PKS genes were introduced into FIESC from other Fusarium lineages. CONCLUSION: Our results suggest that although the portion of the genome devoted to SM biosynthesis has remained similar during the evolutionary diversification of FIESC, the ability to produce SMs could be affected by the different distribution of related functional and complete gene clusters.

Antimicrobial Activity of Yeast Cell Wall Products Against Clostridium perfringens.

Yeast cell wall (YCW) products are used worldwide as alternatives to antibiotics growth promoters for health and performances improvement in livestock. The success of yeast and YCW products as feed additives in farm animals' nutrition relies on their capacity to bind enteropathogenic bacteria and on their immunomodulatory activity. In vivo studies report their anti-infectious activity on Gram-positive pathogens like clostridia. However, the in vitro antimicrobial activity of YCW products seems to be limited to some Gram-negative enteropathogens, and literature lacks in vitro evidences for antimicrobial effect of YCW products against Clostridium perfringens. This study aims to measure the antimicrobial activity of YCW products on C. perfringens. Five different YCW products were assayed for their capacity to inhibit the growth of C. perfringens, by analyzing the growth kinetics of the pathogen. All YCW products inhibited the growth of the pathogen, by reducing the growth rate and the maximum growth value and extending the lag phase duration. The effect on the growth parameters was product and dosage dependent. The most effective YCW (namely YCW2), at the minimum effective concentration of 1.25 mg/mL, increased the lag phase duration by 3.6 h, reduced the maximum growth rate by >50%, and reduced the final cell count by 102 colony-forming unit per milliliter in 24 h, with respect to the control. YCW products did not show a strain-dependent impact on C. perfringens growth when tested on different strains of the bacterium.

Equilibrium Isotherm Approach to Measure the Capability of Yeast Cell Wall to Adsorb Clostridium perfringens.

Yeast cell wall (YCW) products are currently used as substitutes to antibiotic growth promoters, to improve animal performances, and to reduce the incidence of infectious diseases in livestock. They are claimed to bind enteropathogens, thus interfering with their colonization in the intestinal mucosa. Although the anti-infectious activity of YCW products on Gram-positive pathogens like Clostridium perfringens has been reported in vivo, in vitro evidences on the adsorption of C. perfringens by YCW fractions are not yet available. Preliminary results showed that purified YCW products exert antimicrobial activity toward C. perfringens. Using the adsorption isotherm approach, we measured the ability of YCW products in adsorbing C. perfringens, thus reducing its viability. Dosages of YCW products >1 mg/mL adsorbed 4 Log colony-forming unit (CFU)/mL of C. perfringens in buffered solution. The maximum adsorption of the bacterium was reached in 3 h, whereas only one product of four YCW products retained the adsorption up to 6 h. The analysis of equilibrium isotherms and adsorption kinetics revealed that all products adsorb C. perfringens in a dose- and time-dependent manner, with high affinity and capacity, sequestering up to 4 Log CFU/mg of product. The determination of adsorption parameters allows to differentiate among adsorbents and select the most efficient product. This approach discriminated among YCW products more efficiently than the antimicrobial assay. In conclusion, this study suggests that the ability of YCW products in reducing C. perfringens viability can be the result of an adsorption mechanism.

Comparative efficacy of agricultural by-products in sequestering mycotoxins.

BACKGROUND: Biosorption using agricultural by-products has been proven as a low-cost and safe way to sequester mycotoxins. Few agricultural by-products have been studied for their efficacy in adsorbing simultaneously a large range of mycotoxins. The present work compared the ability of 51 agricultural by-products to adsorb mycotoxins from liquid mediums simulating physiological pH values, and it studied the mechanism for mycotoxin adsorption by isotherm adsorption experiments. RESULTS: Grape pomaces, artichoke wastes, and almond hulls were selected as promising biosorbents for mycotoxins, being quite effective towards aflatoxin B1 (AFB1 ), zearalenone (ZEA), and ochratoxin A (OTA). Their adsorption was not affected by medium pH, and the adsorbed fraction was not released when pH rose from acid to neutral values. Fumonisin B1 (FB1 ) was adsorbed to a lesser extent, and deoxynivalenol adsorption was not recorded. For the selected biosorbents, maximum adsorption capacity calculated by the best fitting model (Freundlich, Langmuir, or Sips equation) ranged from 1.2 to 2.9 µg mg-1 for AFB1 , 1.3 to 2.7 µg mg-1 for ZEA, 0.03 from 2.9 µg mg-1 for OTA, and 0.01-1.1 µg mg-1 for FB1 . CONCLUSION: This study confirms that some agricultural by-products can find technological applications as feed/food additives for mycotoxin reduction. © 2018 Society of Chemical Industry.

Efficacy of yeast starters to drive and improve Picual, Manzanilla and Kalamàta table olive fermentation.

BACKGROUND: Table olive fermentation is an unpredictable process and frequently performed using traditional practices often inadequate to obtain products with acceptable quality and safety standards. In the present study, the efficacy of selected yeast strains as starters to drive fermentations of green and black table olives by the Greek method was investigated. Pilot-scale production by spontaneous fermentation as a control, olives started with previously selected Saccharomyces cerevisiae strains and fermentation driven by commercial S. cerevisiae baker's yeast strain were carried out for each of Manzanilla, Picual and Kalamàta table olive cultivars. RESULTS: Time of fermentation was significantly shortened to 40 days to complete the transformation process for all three tested cultivars. Inoculated table olives were enhanced in their organoleptic and nutritional properties in comparison with corresponding samples obtained by spontaneous fermentation. The use of starters was also able to improve safety traits of table olives in terms of biogenic amine reduction as well as absence of undesired microorganisms at the end of the process. CONCLUSIONS: Autochthonous, but also non-autochthonous, yeasts can be used to start and control table olive fermentations and can significantly improve quality and safety aspects of table olives produced by many smallholder farmers. © 2018 Society of Chemical Industry.

Eubiotics for Food Security at Farm Level: Yeast Cell Wall Products and Their Antimicrobial Potential Against Pathogenic Bacteria.

The population increase in the last century was the first cause of the industrialization of animal productions, together with the necessity to satisfy the high food demand and the lack of space and land for the husbandry practices. As a consequence, the farmers moved from extensive to intensive agricultural systems and introduced new practices, such as the administration of antimicrobial drugs. Antibiotics were then used as growth promoters and for disease prevention. The uncontrolled and continuous use of antibiotics contributed to the spread of antibiotic resistance in animals, and this had adverse impacts on human health. This emergence led the European Union, in 2003, to ban the marketing and use of antibiotics as growth promoters, and for prophylaxis purposes from January 2006. This ban caused problems in farms, due to the decrease in animal performances (weight gain, feed conversion ratio, reproduction, etc.), and the rise in the incidence of certain diseases, such as those induced by Clostridium perfringens, Salmonella, Escherichia coli, and Listeria monocytogenes. The economic losses due to the ban increased the interest in researching alternative strategies for the prophylaxis of infectious diseases and for health and growth promotion, such as feed additives. Yeast-based materials, such as cell wall extract, represent promising alternatives to antibiotics, on the base of their prebiotic activity and their claimed capacity to bind enteropathogenic bacteria. Several authors reported examples of the effectiveness of yeast cell wall products in adsorbing bacteria, but there is a lack of knowledge on the mechanisms involved in this interaction. The purpose of this review is to provide an overview of the current approaches used for the control of pathogenic bacteria in feed, with a particular focus on the use of yeast-derived materials proposed to control zoonoses at farm level, and on their effect on animal health.

Artichoke Polyphenols Produce Skin Anti-Age Effects by Improving Endothelial Cell Integrity and Functionality.

Artichoke is a characteristic crop of the Mediterranean area, recognized for its nutritional value and therapeutic properties due to the presence of bioactive components such as polyphenols, inulin, vitamins and minerals. Artichoke is mainly consumed after home and/or industrial processing, and the undersized heads, not suitable for the market, can be used for the recovery of bioactive compounds, such as polyphenols, for cosmetic applications. In this paper, the potential skin anti-age effect of a polyphenolic artichoke extract on endothelial cells was investigated. The methodology used was addressed to evaluate the antioxidant and anti-inflammatory activities and the improvement of gene expression of some youth markers. The results showed that the artichoke extract was constituted by 87% of chlorogenic, 3,5-O-dicaffeoylquinic, and 1,5-O-dicaffeoylquinic acids. The extract induced important molecular markers responsible for the microcirculation and vasodilatation of endothelial cells, acted as a potential anti-inflammatory agent, protected the lymphatic vessels from oxidative damage by ROS formation, and enhanced the cellular cohesion by reinforcing the tight junction complex. In addition, the artichoke extract, through the modulation of molecular pathways, improved the expression of genes involved in anti-ageing mechanisms. Finally, clinical testing on human subjects highlighted the enhancement by 19.74% of roughness and 11.45% of elasticity from using an artichoke extract cosmetic formulation compared to placebo cream.

A new solid-phase extraction and HPLC method for determination of patulin in apple products and hawthorn juice in China.

A new solid-phase extraction (SPE) pretreatment method using a home-made polyvinylpolypyrrolidone-florisil (PVPP-F) column was developed for the analysis of patulin in apple and hawthorn products in China. Fifty samples (25 apple juices, 12 apple jams, and 13 hawthorn juices) were prepared using the new method and then analyzed by high performance liquid chromatography with diode array detection (HPLC-DAD) on an Agela Venusil MP C(18) reversed-phase column (4.6 mm × 250 mm, 5 µm). The cleanup results for all samples using home-made PVPP-F column were compared with those obtained using a MycoSep®228 AflaPat column. The correlation coefficient R (0.9998) fulfilled the requirement of linearity for patulin in the concentration range of 2.5-250 µg/kg. The limits of detection (LODs) and quantification (LOQs) of patulin were 3.99 and 9.64 µg/kg for PVPP-F column, and 3.56 and 8.07 µg/kg for MycoSep®228 AflaPat column, respectively. Samples were spiked with patulin at levels ranging from 25 to 250 µg/kg, and recoveries using PVPP-F and MycoSep®228 AflaPat columns were in the range of 81.9-100.9% and 86.4-103.9%, respectively. Naturally occurring patulin was found in 2 of 25 apple juice samples (8.0%) and 1 of 13 hawthorn juice samples (7.7%) at concentrations ranging from 12.26 to 36.81 µg/kg. The positive results were further confirmed by liquid chromatography electrospray ionization mass spectrometry (LC-ESI-MS).

foodsafety4eu: paving the way for the food safety system of the future.

Food safety is a top priority for the European Commission, which policies aim at sustaining a high level of protection of human health and consumers' interests, while ensuring an effective functioning of the internal EU market. Under the New Transparency Regulation (EU/2019/138), the 3-year EU-funded foodsafety4eu project (FS4EU) kicked off in January 2021, represents a significant step for the European Union (EU) food safety system (FSS), towards more transparency, better engagement, and closer cooperation. This Horizon 2020 Project, coordinated by CNR-ISPA (Italy), focuses on building a multi-stakeholder platform for the future EU FSS. The foodsafety4eu Network currently consists of 23 consortium partners and around 50 stakeholders: Food Safety Authorities (FSA), consumer associations, academia, research centres and networks, food industries and sector associations, thinktanks, etc. Through a structured, digitally supported, participatory process, the platform hosts the co-design of future strategic research and innovation agenda (SRIA), as well as risk communication models tailored to the specificities of various target groups. Among the goals: providing scientific advice and technical support for EU food safety policies, by enabling actors to access, share and exchange scientific knowledge, resources, and data more efficiently, to better synchronise food safety research and policies, and to contribute to a more transparent communication through the FSS. Overall, the FS4EU project underpins the EFSA missions in risk assessment, including risk communication, with the ambition to be a basis for a Knowledge Centre for Food Safety in Europe. Accordingly, the EU-FORA 2021-2022 fellow (based in CNR-ISPA under the Work Programme RECIPE: Risk AssEssment/Risk CommunIcation: understanding the context and addressing Priorities of the futurE - a learning-by-doing approach) committed to multiple Work Packages, actively participated to various activities, thus successfully contributing to the FS4EU project meeting its objectives by the end of 2023.

Simultaneous Removal of Mycotoxins by a New Feed Additive Containing a Tri-Octahedral Smectite Mixed with Lignocellulose.

Simultaneous removal of mycotoxins has been poorly addressed, and a limited number of studies have reported the efficacy of feed additives in sequestering a large spectrum of mycotoxins. In this study, a new mycotoxin-adsorbing agent was obtained by properly mixing a tri-octahedral smectite with a lignocellulose-based material. At a dosage of 1 mg mL-1, these materials simultaneously adsorbed frequently occurring mycotoxins and did not exert a cytotoxic effect on intestinal cells. Chyme samples obtained by a simulated GI digestion did not affect the viability of Caco-2TC7 cells as measured by the MTT test. In addition, the chyme of the lignocellulose showed a high content of polyphenols (210 mg mL-1 catechin equivalent) and good antioxidant activity. The properties of the individual constituents were maintained in the final composite, and were unaffected by their combination. When tested with a pool of seven mycotoxins at 1 µg mL-1 each and pH 5, the composite (5 mg mL-1) simultaneously sequestered AFB1 (95%), FB1 (99%), ZEA (93%), OTA (80%), T-2 (63%), and DON (22%). HT-2 adsorption did not occur. Mycotoxin adsorption increased exponentially as dosage increased, and occurred at physiological pH values. AFB1, ZEA and T-2 adsorption was not affected by pH in the range 3-9, whereas OTA and FB1 were adsorbed at pH values of 3-5. The adsorbed amount of AFB1, ZEA and T-2 was not released when pH rose from 3 to 7. FB1 and OTA desorption was less than 38%. Langmuir adsorption isotherms revealed high capacity and affinity for adsorption of the target mycotoxins. Results of this study are promising and show the potential of the new composite to remove mycotoxins in practical scenarios where several mycotoxins can co-occur.