Giuseppe Meca

Contamination of food and feed by aflatoxin B1 (AFB1) remains a major global concern due to its toxicity, carcinogenicity, and persistence in the food chain. Environmental and climatic pressures continue to favor aflatoxigenic fungal contamination, highlighting the need for effective and sustainable detoxification strategies. Biological detoxification has emerged as a promising alternative to conventional physical and chemical treatments. This systematic review, conducted following PRISMA guidelines, summarizes microbial and enzymatic approaches for AFB1 detoxification, focusing on bacteria, yeasts, and microbial enzymes. The literature shows a predominance of bacterial systems, especially lactic acid bacteria and Bacillus species, mainly acting through adsorption, fungal growth inhibition, and suppression of aflatoxin biosynthesis. Yeasts, although less represented, also showed promising detoxification capacities through adsorption and biodegradation-related mechanisms. Enzymatic systems achieved the highest efficiencies, particularly oxidative enzymes such as laccases and dye-decolorizing peroxidases, often exceeding 90% detoxification under optimized conditions. However, industrial application remains limited by laboratory-scale validation, variability among protocols, incomplete toxicological assessment of degradation products, and limited evidence in complex food and feed matrices.

Vitis vinifera L. leaves represent one of the most phenolic-rich grapevine biomasses, exhibiting a broad spectrum of biological activities, including prebiotic and microbiota-modulating ones. However, as they are mainly regarded as agro-industrial waste, they are still an underexploited resource. Probiotic-driven fermentation may offer a promising strategy to enhance their bioactive potential while addressing this challenge. The aim of this study was to evaluate the effects of the in vitro fermentation of Vitis vinifera L. leaf extract (VVLE) by Lactiplantibacillus plantarum PBS067 in terms of phenolic content, antioxidant activity, lactic acid and protein content, and metabolomic profile. Our analysis shows the prebiotic activity of VVLE, as VVLE-supplemented medium allowed probiotic growth without toxic effects. Leaves fermentation led to a significant enrichment in lactic acid and the biotransformation of complex plant-secondary metabolites into smaller derivatives. However, VVLE antioxidant capacity slightly decreased after the fermentation, likely due to the phenolic conversion itself. These results highlight how controlled lacto-fermentation valorizes grapevine leaves, transforming them into sustainable upcycled bioactive products with potential applications within cosmetics, pharmaceuticals, nutraceuticals and complementary medical product sectors. Key points: • L. plantarum PBS067 thrives in medium supplemented with polyphenol-rich leaf extract • L. plantarum PBS067-driven fermentation modulates VVLE phytochemical profile • L. plantarum PBS067-driven fermentation valorizes viticulture by-products

Reusing waste materials is a sustainable practice to reduce the impact of human activities on the global ecosystem. In particular, agro-industrial waste can be explored as an innovative and green source of beneficial bioactive compounds. For this purpose, seventeen different varieties of wine grape pomace were collected from the Apulia region (Italy) to perform extractions and qualitative–quantitative phenolic profile characterization. To ensure an environmentally friendly extraction process, natural deep eutectic solvents (NADESs) were used as privileged media. After an initial screening, Merlot (MEA) and Sangiovese (SGA) varieties were then selected for extraction with betaine/lactic acid 1:4 (mol/mol) +40% water (w/w) and choline chloride/ascorbic acid 2:1 (mol/mol) +40% water (w/w). They were further investigated by LC-QTOF-MS for a comprehensive metabolomic evaluation. Betaine-based extracts from both cultivars exhibited higher polyphenol contents than choline chloride-based ones: 175.0 and 161.8 mg/kg compared to 59.6 and 40.4 mg/kg. Qualitative antimicrobial assays confirmed the effectiveness of betaine/lactic acid (MEA) and choline chloride/ascorbic acid (SGA) extracts on B. allii and E. coli, respectively, as well as the NADES themselves, even though it is difficult to discriminate the individual effects. Finally, the evaluation of both antioxidant activity and total polyphenol content led to good results for betaine/lactic acid (2.8 and 3.2 mg TEAC/g DW and 0.92 and 0.93 mg GAE/g DW for SGA and MEA, respectively), while the content of ascorbic acid, used as a component of NADES, substantially influenced the relevant obtained results in choline chloride/ascorbic acid samples. These findings support the potential of combining NADES with polyphenols recovered from grape pomace as a promising approach for further optimization and application-oriented investigation.

Durum wheat is increasingly threatened by Fusarium diseases, including Fusarium Crown Rot (FCR) and Fusarium Root Rot (FRR), caused primarily by Fusarium culmorum, F. graminearum, and F. pseudograminearum. These pathogens reduce yield, by causing pre- and post-emergence damping-off and induce symptoms including stem and root browning, as well as severe growth stunting. Traditional control methods, including agronomic practices and seed treatments with fungicides, face limitations due to pathogen resistance and environmental concerns, emphasizing the need for sustainable, microbiologically-driven alternatives, particularly in organic farming. This study evaluates beneficial bacteria as biocontrol agents for efficacy in controlling Fusarium disease in durum wheat. An in vitro screening of 89 bacterial strains against a panel of Fusarium isolates, specifically Fusarium culmorum (Fc383, Fc401, Fc820, Fc1126, Fc1435, Fc1674), F. graminearum (Fg566, Fg913, Fg919), and F. pseudograminearum (Fp1139, Fp1140), identified promising candidates, including Bacillus, Pseudomonas, and lactic acid bacteria (LAB). Seed biopriming with Bacillus amyloliquefaciens MA8 and two strains of Lactiplantibacillus plantarum (5H1 and HB23) effectively suppressed Fusarium infection in vitro. Lp. plantarum HB23 demonstrated in vivo the highest biocontrol efficacy against Fc1126 and Fc566 strains, reducing disease severity by 44.4% and 49.2%, respectively. Metabolomic profiling revealed bacterial production of antifungal compounds, including surfactin and fengycin from Bacillus and benzoic and hydrocinnamic acids from Lactobacillaceae. This study demonstrates Lp. plantarum as an effective biopriming agent against F. culmorum and F. graminearum in durum wheat, confirming that seed treatment with BCAs is a viable alternative to chemical fungicides, especially in organic agriculture.

Biologically based detoxification strategies are increasingly being explored as alternatives to conventional methods for the removal of toxic contaminants in food products. Among these, aflatoxin B1 (AFB1) is one of the most potent mycotoxins due to its high toxicity, genotoxicity, and persistence in the human body once ingested. In this study, the detoxification potential of bacterial strains belonging to the genera Lactobacillus/Pediococcus (n = 10) and Bacillus (n = 10) was evaluated using extracts from naturally contaminated corn flour. Detoxification was assessed after incubation for 12, 24, and 48 h in specific culture media. AFB1 quantification and metabolite profiling were performed at each time point using Quadrupole Time-of-Flight Mass Spectrometry (LC-QTOF-MS). The highest detoxification rates were observed with Lactobacillus curvatus 14 (L. curvatus 14) (41.1 ± 19.3%) and Pediococcus pentosaceus 4 (P. pentosaceus 4) (25.4 ± 11.3%) after 48 h, and Bacillus firmus 6 (B. firmus 6) (25.1 ± 12.9%) after 24 h. An in vitro digestion model was also applied to assess detoxification under gastrointestinal conditions. Results showed substantial AFB1 reduction at the colonic stage, reaching 72.26 ± 7.54% for P. pentosaceus 4 and 69.67 ± 9.70% for L. curvatus 14. These findings underscore the potential application of Lactobacillus, Pediococcus, and Bacillus strains in biological detoxification strategies to reduce dietary exposure to AFB1.

The rising demand for vegan products calls for new plant-based antimicrobial preservation methods. This study evaluates an antifungal ingredient obtained by fermenting oat drink with lactic acid bacteria to extend vegan mortadella’s shelf life. In vitro tests showed antimicrobial effects against Aspergillus flavus, Penicillium commune, and Listeria monocytogenes (inhibition zones: 2–5 mm). The enrichment of the oat drink culture medium with additional nutrients enhanced fermentation performance and increased antifungal activity. The fermented culture medium with the highest antimicrobial activity was used to develop a bioactive ingredient for the preservation of vegan mortadella conservation. Adding 3% of this ingredient to vegan mortadella improved microbial stability, reducing mesophilic bacteria by 2.5 Log₁₀ CFU/g and increasing lactic acid bacteria. Lower pH and water activity changes were observed but remained within quality standards. Contamination assays showed a consistent reduction of A. flavus over 7 days, while P. commune and L. monocytogenes dropped below detection within 2 days. In contrast, control samples maintained contamination levels near 3.0 Log₁₀ CFU/g. These findings support the potential of fermented oat-based ingredients as effective, natural preservatives for vegan foods.

Consumer demand for natural, additive-free foods is increasing. Following the trend, this study evaluated the antifungal potential of lactic acid bacteria (LAB) against Penicillium species commonly found in cheese, using both LAB ferments and hydroxypropylmethylcellulose (HPMC) coatings. LAB strains were first screened with a dual-culture assay. Fermentations in Man–Rogosa–Sharpe (MRS) broth and milk whey were lyophilized and tested, with whey-based ferments showing greater antifungal activity. All whey ferments inhibited fungal growth, with KK₁₃, KB₂, KB₃, and KB₄ being the most effective based on MIC and MFC assays. KB₃-fermented whey had the highest levels of antifungal metabolites, such as phenyllactic acid. A coating containing 5% HPMC and 100 g/L of KB₃-fermented whey was applied to cheese slices, reducing the fungal counts of Penicillium commune by more than 1 Log₁₀ CFU per gram and extending shelf life by 12 days. In whole-cheese trials with natural contamination, this coating delayed visible fungal growth until day 60, extending shelf life by 45 days compared with uncoated samples and 33 days compared with coated controls. These findings support the use of LAB-fermented whey and HPMC coatings as natural preservation strategies, thereby contributing to the sustainable reuse of dairy by-products.

Zearalenone (ZEA) is a mycotoxin produced by Fusarium species, and cause contamination of food and feed, with impacts in animal production and in food production chain. Effective detoxifying methods, such as biodegradation, are therefore required. This study aimed to isolate microorganisms and screen ZEA detoxifying strains. As a result, 197 microorganisms were isolated, and six were initially selected after colorimetric screening. ZEA (1 µg/mL) was added to culture media, and after 24 h, all six microorganisms were able to degrade ZEA, without the formation of α-ZOL. One isolate eliminated ~ 99% of ZEA and was identified as Bacillus velezensis CL197. ZEA metabolites produced by the bacteria were evaluated, and no metabolites with greater or similar toxicity than ZEA were detected. This strain was applied to swine in vitro digestion, and up to 64% of ZEA was degraded. B. velezensis CL197 significantly degraded ZEA, demonstrating potential to be used as a detoxifying agent in the food production chain as a biocontrol agent.