Knowledge of the ecology of biological control agents (BCAs), previously reported to be effective against the citrus vascular pathogen Plenodomus tracheiphilus, is required to improve scheduling of BCA field applications. Culture-dependent methods (dilution plate and direct plating) and culture-independent qPCR assays were used to determine survival of Bacillus amyloliquefaciens QST 713 and Trichoderma asperellum ICC 012 + T. gamsii ICC 080 on citrus stem and leaf tissues following foliar applications, and root endospheres and rhizospheres following root drenches with these Trichoderma BCAs. Viable population levels of B. amyloliquefaciens did not change over time on treated stems, whereas these decreased in leaf samples after 14 d. The qPCR assay detected B. amyloliquefaciens in all collected samples, with no temporal changes in population levels. Although the qPCR assay detected T. asperellum and T. gamsii in leaf and stem tissues, these fungi were isolated only from stem tissues, with increased isolations at 21 d post treatment compared with 7 d. Neither qPCR nor culturing detected Trichoderma species in citrus vascular root tissues. However, qPCR and culturing detected these fungi in host rhizospheres at 7, 14 and 21 d post inoculation, confirming their rhizosphere competence. This study has provided insights into colonization and survival within citrus plants of B. amyloliquefaciens and T. asperellum + T. gamsii contained in commercial biocontrol products. These indicate that integrating qPCR and culture-dependent approaches is important for detecting and quantifying these BCAs. The endophytic lifestyle of B. amyloliquefaciens and T. asperellum + T. gamsii makes them likely to provide long-term biological control. These results also indicate that the selected Bacillus and Trichoderma agents could spread and colonize citrus tissues over extended periods, and especially after host pruning or damage, which could promote plant colonization and protection from pathogens.

This study aimed to explore the culturable bacterial community in healthy volunteers, with a focus on the staphylococcal population. First, 120 bacterial isolates were collected from conjunctival swabs of 49 healthy volunteers (aged 18 to 77 years). Isolates were characterized by phenotypic analyses, and 68 of them were confirmed as staphylococci by MALDI-TOF/MS and a multiplex PCR assay. The Staphylococcus strains (n = 68) were tested for resistance to the most relevant clinical antibiotics. Clonal relationships among isolates were evaluated by pulsed-field gel electrophoresis (PFGE), and the presence of mecA and mecC, as well as pvl genes, was investigated. Staphylococcus aureus (74

The growing demand for natural and sustainable food preservation strategies has driven the development of innovative edible biopackaging solutions. This study aimed to formulate and evaluate the role of two novel anti-Listeria edible bioactive packaging solutions, based on whey protein (WPS) and tapioca starch (TSS), both activated with the commercial bacteriophage Listex™ P100. Physical analyses revealed that WPS exhibited higher density (1.08 g/mL), lower surface tension (56.9 mN/m), and a higher negative zeta potential (-37.0 mV) compared to TSS (0.97 g/mL, 74.7 mN/m, -4.4 mV, respectively), indicating superior spreadability and electrostatic stability than TSS packaging. WPS films also demonstrated greater thickness (0.188 mm vs. 0.094 mm) and lower transparency (759.13 % vs. 233.56 %) than TSS films. Both formulations maintained microbiological safety and phage viability for up to 60 days under refrigerated conditions. In vivo trials, on artificially contaminated cheese samples, showed a significant reduction in Listeria monocytogenes counts [≈1.1 log colony forming units (CFU)/g] after 3 days of storage when treated with either WPS or TSS phage containing coatings, compared to untreated controls. From a sensory standpoint, mini cheeses coated with WPS were rated as having excellent overall quality, characterized by a typical appearance and absence of visual defects. These findings provide a preliminary proof-of-concept for the use of phage-activated edible coatings as a sustainable and safe strategy to improve cheese safety.

Lactic acid bacteria (LAB) are widely used in food systems; among them, bacteriocin-producing strains have attracted attention for their potential in the biopreservation of dairy products. This study started from the detection of bacteriocin-encoding genes in eight probiotic Leuconostoc mesenteroides subsp. mesenteroides strains, previously isolated, identified, and characterized for antimicrobial activity. Results confirmed the presence of bacteriocin genes across the strains, with Ln.F5 harboring both mesB and lcnA genes, and three other strains, including the Ln.M14 strain, exclusively carrying the lcnA gene. The two strains, Ln.F5 and Ln.M14, were used, in single and mixed cultures, for the first time, as adjunct cultures in a model cheese. Their impact against Listeria spp., Staphylococcus aureus, Escherichia coli, Micrococcus luteus, and Brochothrix thermosphacta, and on volatile organic compounds (VOCs), during ripening and storage, was evaluated. Results showed high viability (9.2 Log CFU/g) of Leuconostoc spp. in model cheese, up to 60 days of storage, and Pulsed-Field Gel Electrophoresis (PFGE) profiles of the re-isolated bacteria confirmed the survival of the added strains. Furthermore, results indicated the inhibition of E. coli and Listeria spp. started from the 15th day of ripening in samples differently inoculated with the two Leuconostoc strains. Listeria spp. was completely inhibited starting from 15 days by Ln.M14, in single culture. The complete inhibition of S. aureus, M. luteus, and B. thermosphacta was detected after 30 days of ripening in samples differently inoculated with Ln.F5 and Ln.M14. The VOC analyses revealed more complex aromatic profiles in samples inoculated with Leuconostoc strains, which, along with the development of cheese eyes, confirmed the effect of the Leuconostoc strains in enhancing quality traits of cheeses.

Minas artisanal cheese (MAC) is a traditional raw-milk cheese from Minas Gerais, Brazil, whose safety depends on hygienic-sanitary controls and artisanal practices. This study evaluated the microbiological quality of MAC according to Minas Gerais state criteria and investigated associations with production characteristics. Ninety-six cheese samples from 32 producers were analyzed for aerobic mesophiles, Enterobacteriaceae, coliforms, Escherichia coli, coagulase-positive staphylococci (CPS), enterotoxins, Listeria monocytogenes, and Salmonella spp. Non-compliance was observed in 39.6% of samples for coliforms, 6.3% for E. coli, and 21.9% for CPS; overall, 53.1% failed at least one criterion. Coliform compliance was associated with producing region (p < 0.05), whereas no regional association was observed for E. coli or CPS. L. monocytogenes, Salmonella spp., and enterotoxins were not detected. These findings indicate regional variation in hygiene conditions and support region-specific interventions to improve MAC microbiological quality.

This study aimed to evaluate the antimicrobial and antibiofilm potential of 15 LAB strains using five types of LAB-derived preparations: cell-free supernatants (CFS), sonicated-inactivated cells (IC), their combination (ICS), and their neutralized variants (CFS N, ICS N) to identify the most effective strain-extract combinations for potential application as natural biocontrol agents in dairy systems. Antimicrobial activity was assessed through agar diffusion assays, growth and biofilm inhibition, and determination of minimum inhibitory (MIC) and bactericidal (MBC) concentrations. The extracts were further characterized by pH and organic acid profiles using high-performance liquid chromatography (HPLC), and their mechanisms of action were investigated through cellular leakage assays, time-kill kinetics, and scanning electron microscopy (SEM). Several LAB-derived extracts exhibited strong antagonistic and antibiofilm activity against both spoilage and pathogenic bacteria. Non-neutralized CFS and ICS showed pronounced bactericidal activity, confirming the central role of organic acids in microbial inhibition. Z-score ranking and leakage assays identified Lactiplantibacillus plantarum Q4C3, Lactococcus lactis subsp. lactis biovar diacetylactis SBR4, Weissella cibaria W21, and Weissella viridescens W23 as the most effective strains. Time-kill assays demonstrated rapid microbial reductions (>3 log CFU/mL) within 4 h by CFS, whereas ICS required longer exposure. SEM analysis revealed severe membrane disruption in CFS-treated cells and the presence of LAB-derived debris surrounding ICS-exposed cells. These findings demonstrate that acidic LAB-derived extracts, particularly CFS, efficiently disrupt microbial cells and support their use as safe and effective natural biocontrol agents for improving the microbial safety and quality of dairy products.

Mediterranean buffalo meat is a nutrient-rich, low-fat resource with high protein and favorable lipid profiles. Its use in fermented products, like buffalo salami, remains still limited, despite its economic and cultural importance in regional livestock systems. This study evaluated the impact of four treatments on microbiological succession, lipid composition, volatile compounds, and sensory properties of buffalo salami. The treatments were: not inoculated control (CTR), commercial starter culture (S1), commercial starters combined with Lacticaseibacillus rhamnosus E26 and Lactiplantibacillus plantarum M5.1 strains (S2), and a combination of L. rhamnosus E26 and L.plantarum M5.1 strains (S3). Results showed that all treatments did not alter nutritional composition but significantly enhanced lipid quality. S2 and S3 samples reduced cholesterol levels compared to the inoculated control and increased polyunsaturated fatty acids. Atherogenic and thrombogenic indices were lower S1 samples, suggesting a potentially more favorable lipid profile from a nutritional perspective. Microbiological analyses confirmed the absence of Salmonella spp. and Listeria monocytogenes, a drastic reduction of staphylococci, after 40 days of ripening, and a quite stable level of lactic acid bacteria. Volatile profiles revealed modulation of acids, aldehydes, ketones, and terpenes, consistent with sensory outcomes, with S2 and S3 samples having the highest acceptability, improved aroma and texture. Collectively, targeted use of L. plantarum and L. rhamnosus strains, as starter cultures, improved microbiological safety, influenced lipid-related nutritional parameters, and improved sensory characteristics, highlighting their potential for the development of high-value buffalo salami. These findings underscore the strategic role of lactic acid bacteria in optimizing functional and organoleptic properties of fermented meat products.

Background/Objectives: In food-producing animal (FPA) environments, healthy animals can act as reservoirs of potentially pathogenic Escherichia coli, which can be transmitted through the food chain to humans. This study aimed to evaluate cloacal E. coli in healthy Sicilian lambs subjected to an experimental feeding regimen by assessing bacterial levels, antimicrobial resistance, virulence traits, and the clonal relationships, as well as the impact of a pistachio skin as an agro-industrial by-product supplement during a 58-day feeding trial. Methods: A total of 295 E. coli isolates from the control (CTRL) and treatment (Treated) groups at initial time (T0) and final time (T1) were phenotypically and genotypically characterized using Kirby-Bauer antimicrobial testing, multiplex PCR for virulence genes, and PFGE for clonal analysis. Results: The feeding regimen did not significantly influence the prevalence, abundance, or virulence of the E. coli isolates. Shiga toxin-producing E. coli (STEC) were the most common pathotype, mainly carrying the stx1 gene, while the Enteroinvasive (EIEC) type was detected only sporadically. Enteropathogenic E. coli (EPEC) predominated at T0, while enteroaggregative E. coli (EAEC) at T1, and enterotoxigenic E. coli (ETEC), initially prevalent in Treated samples, disappeared by T1. Antimicrobial resistance profiles varied among isolates, with the highest resistance observed in the CTRL group. However, both groups exhibited high resistance to streptomycin, and 9% of CTRL isolates were multidrug resistant. A notable reduction in overall resistance rates, especially in the Treated group, was observed, indicating a dietary effect on the E. coli resistome. PFGE genotyping showed high genetic diversity, with resistance traits more frequently detected than virulence factors. Conclusions: This study highlights that healthy lambs serve as reservoirs for potentially human-pathogenic E. coli and suggests that dietary regimes could effectively reduce antibiotic resistance.

This study presents the first comprehensive assessment of a bacteriophage P100-activated edible whey-protein solution (WPS) applied to the rind of Sicilian Canestrato Fresco (SCF) cheese. Beyond evaluating its anti-Listeria efficacy in pre- and post-packaging contamination contexts, the work investigates the coating’s effects on chemical composition, volatilome, sensory properties, and consumer responses, including willingness to pay. To assess anti-Listeria activity, all samples were stored at 4 °C for 30 days. Contamination was carried out either before or after coating application, depending on the specific treatment. Listeria monocytogenes was monitored at 0, 1, 3, 7, 15, and 30 days of refrigerated storage. The active coating reduced the pathogen from approximately 3 log CFU/g to undetectable levels (0 log CFU/g) within 3 days, whereas the untreated controls reached about 5 log CFU/g after 30 days. WPS-coated cheeses showed no significant changes in chemical composition (moisture ~33%, protein ~29%, fat ~33%) or fatty acid profile compared to traditional SCF. The volatilome was dominated by hexanoic and butanoic acids and ethyl esters, without significant differences between coated and control samples, as confirmed by Smart Nose® analysis. Sensory evaluation by trained assessors demonstrated that the bioactive coating did not alter the traditional sensory profile of SCF cheese. A consumer survey conducted with 240 participants from two retail formats revealed significant differences in product familiarity and perceived food safety, while openness to innovation and willingness to pay were similar. More than 90% of respondents were willing to pay a 10% price premium. Overall, phage-based edible coatings appear to be edible, renewable, and biodegradable packaging alternative to improve cheese safety without compromising quality.

Germinated brown rice (GBR) has gained considerable attention as a functional food due to both nutritional and bioactive profiles as well as health-promoting properties. The present narrative review is aimed at summarizing and discussing available research on GBR, focusing on bioactive composition and potential to support human well-being. Based on available data, the germination process of brown rice enhances the bioavailability of key bioactive compounds, including polyphenols, γ-aminobutyric acid, γ-oryzanol, vitamins, and dietary fibers. Additionally, a substantial body of evidence supports the ability of GBR to exert beneficial physiological effects on the host, such as modulation of lipid and glucose metabolism, antioxidant and anti-inflammatory activities, and positive influences on gut microbiota composition. The use of GBR in formulating functional foods and nutraceutical supplements further highlights its versatility as a promising strategy in supporting human well-being.

This study evaluated the direct use of lactose, the main sugar fraction of dairy side-streams, for the mixotrophic cultivation of Tetraselmis chuii. An equimolar glucose/galactose mixture, representing the pre-hydrolysed equivalent of lactose, was included as a comparative treatment. Batch cultures were grown in synthetic media for 10 days at three carbon levels (2.0, 3.5, and 5.0 % w/v), using a photoautotrophic culture as a control. Growth performance, nutrient removal, biochemical productivities, and cell adhesion were assessed. The 3.5 % lactose level showed the best overall performance, achieving a specific growth rate of 0.49 day-1, a biomass productivity of 1.26 × 105 cells mL-1 day-1, and a maximum cell density of 1.50 × 106 cells mL-1. It also supported a balanced biomass composition, with protein, carbohydrate, fatty acid, and carotenoid productivities of 213, 154.71, 57.29, and 2.63 mg L-1 day-1, respectively. In addition, under mixotrophic cultivation, PEG/PDMS coating reduced cell adhesion by approximately 50 % compared with glass. These results highlight the potential of direct lactose use as a carbon source for the mixotrophic cultivation of T. chuii, supporting a sustainable strategy for the valorization of lactose-rich dairy by-products.

The dairy industry has increasingly seeking natural strategies to enhance food safety and quality. Fungal contamination remains a major challenge due to spoilage, economic losses, and mycotoxin production, making bioprotective microbial cultures a promising alternative. This study aimed to investigate the antifungal potential of five Lactiplantibacillus plantarum strains (M3.1, M3.3, M3.6, R3.2, and R3.6) against Aspergillus niger and Penicillium chrysogenum. All strains significantly inhibited fungal growth, as demonstrated by in vitro assays and reductions in fungal biomass. Scanning electron microscopy (SEM) revealed severe morphological damage to fungal spores, including surface disruption, deformation, and shrinkage. Metabolomic analyses identified the production of organic acids, including lactic, acetic, succinic, malic, propionic, butyric, and formic acids, using high-performance liquid chromatography (HPLC), as well as 35 additional metabolites, mainly amino acids, fatty acids, cyclic dipeptides, phenolic compounds, and esters, using high-performance liquid chromatography-mass spectrometry (HPLC-MS), revealing a synergistic and multifactorial antifungal mechanism. In a cheese model system, all strains completely inhibited P. chrysogenum, while strains M3.3, M3.6, and R3.2 fully inhibited A. niger. These findings demonstrate the strong antifungal activity of L. plantarum strains and support their application as bioprotective cultures in dairy products.

The edible insect market has grown significantly due to their high nutritional value, digestibility, palatability, and functional properties. They are straightforward to cultivate and present a low environmental impact, keeping a high protein content regardless of diet. In this study, Tenebrio guineensis larvae were fed a mixture of various local food by-products (FBPs) at 33.3

Pistachio skin is a by-product that is considered a promising novel feed ingredient for ruminants; however, its role in shaping the lamb gastrointestinal tract microbiota is poorly studied. The present study aimed to investigate, through a metagenomics approach, the effects of integrating pistachio skin into the diet on the faecal and ruminal microbiota of healthy lambs. Faecal samples, collected at the beginning (d0) and 58 days after the start of the dietary treatment (d58), and ruminal samples, collected after slaughter, were subjected to Illumina MiSeq analysis of the 16S rRNA gene. The results revealed that, although temporal variations were observed, the supplementation of pistachio skin did not markedly affect the overall faecal microbiota structure. Conversely, specific rumen taxa were selectively modulated by the experimental diet. In conclusion, the use of pistachio skin as a feed ingredient can be considered a suitable and sustainable dietary strategy that modulates specific rumen microbial groups, thereby preserving the stability of the gut microbiota in lambs.

The dairy sector produces considerable amounts of nutrient-rich effluents, which are frequently undervalued as simple by-products or waste. In particular, Second Cheese Whey (SCW), also known as scotta, exhausted whey, or deproteinized whey, represents the liquid fraction from ricotta cheese production. Despite its abundance and high organic and saline content, SCW is often improperly discharged into terrestrial and aquatic ecosystems, causing both environmental impact and resource waste. The available purification methods are expensive for dairy companies, and, at best, SCW is reused as feed or fertilizer. In recent years, increasing awareness of sustainability and circular economy principles has increased interest in the valorization of SCW. Biological treatment of SCW using microalgae represents an attractive strategy, as it simultaneously reduces the organic load and converts waste into algal biomass. This biomass can be further valorized as a source of proteins, pigments, and bioactive compounds with industrial relevance, supporting applications in food, nutraceuticals, biofuels, and cosmetics. This review, starting from analyzing the characteristics, production volumes, and environmental issues associated with SCW, focused on the potential of microalgae application for their valorization. In addition, the broader regulatory and sustainability aspects related to biomass utilization and treated SCW are considered, highlighting both the promises and limitations of microalgae-based strategies by integrating technological prospects with policy considerations.

The Etna region has recently emerged as a producer of excellent wine, and autochthonous yeasts are gaining recognition for enhancing wine's aromatic complexity.The present study aimed at evaluating the effect of indigenous yeasts, such as Hanseniaspora uvarum and Starmerella bacillaris, used in sequential fermentation with Saccharomyces cerevisiae, on the Carricante grape variety through the evaluation of sugars, target organic acids, ethanol content, aromatic compounds, and sensory attributes. Results indicated that the alcoholic fermentation with St. bacillaris or H. uvarum and native S. cerevisiae showed the fastest kinetics, completing within 12 days, in samples inoculated with H. uvarum and the commercial S. cerevisiae strain it took 14 days, whereas must inoculated with St. bacillaris and commercial S. cerevisiae showed a slight slower fermentation. The volatile profiles included 55 compounds, mainly alcohol and esters. Phenylethyl alcohol was significantly higher in wines fermented with St. bacillaris, wines fermented with H. uvarum showed higher ester content. These distinct metabolic contributions underline the complementary roles of yeasts in promoting the expression of terroir and shaping wines with unique aromatic profiles. Moreover, wines fermented with H. uvarum exhibited higher sensory satisfaction, confirming that fermentation strategies can affect volatile compound profiles and enhance both sensory quality and market acceptance.

This study aimed to assess the technological, safety, antibiotic resistance, and antifungal properties of 35 lactic acid bacteria (LAB) from Sicilian dairy products. Sixteen Lactiplantibacillus plantarum isolates produced diacetyl, and 30 LAB isolates showed proteolytic activity, suggesting technological potential. All isolates presented weak milk acidification capacity. Aminopeptidase activity varied, with PepX (5/35) and PepN (2/35) showing better activity on Lactiplantibacillus fermentum, Lactobacillus sp. and L. plantarum strains, indicating species-specificity. None of the isolates showed hemolytic, gelatinase, DNase, or coagulase activity. Most were resistant to oxacillin, tetracycline, rifampicin, and vancomycin. Regarding antifungal activity, L. plantarum (n = 10) and Lactiplantibacillus sp. (n = 1) isolates inhibited Aspergillus niger and Penicillium chrysogenum, while L. rhamnosus (n = 5) strains were active against P. chrysogenum. Overall, Lactiplantibacillus isolates, particularly L. plantarum from donkey milk, showed promising technological traits, safety, and potential as bioprotective cultures. These findings support the search for new strains with improved characteristics and technological applications.

The aim of the present study was to develop a synbiotic formulation based on germinated brown rice (GBR) and probiotics with health-promoting benefits. The GBR was selected based on its well-known role as a prebiotic substrate rich in bioactive compounds, particularly gamma-oryzanol and gamma-aminobutyric acid, able to exert antioxidant and anti-inflammatory activities. Four probiotic strains (Lacticaseibacillus rhamnosus CA15, Limosilactibacillus fermentum CS57, Lactiplantibacillus plantarum IMC510, and Bifidobacterium animalis subsp. lactis BLC1) were selected and in vitro tested, single and as a blend, for the ability to survive in the simulated gastrointestinal tract. Simultaneously, the GBR was investigated for antioxidant, anti-inflammatory, glycogen release, and cell proliferation abilities. Synbiotic formulations, based on a blend of the four selected probiotics and GBR at three different concentrations (GBR1, 40 %; GBR2, 30 %; GBR3, 20 %), were set up to evaluate the survivability of the probiotics and the antagonistic activity against pathogens. Results revealed a good survivability of the probiotic strains under in vitro simulated gastrointestinal transit and in presence of different GBR concentrations. The synbiotic formulation containing the lowest GBR concentration (GBR3) showed the broadest spectrum of antagonistic activity against the tested pathogens. The GBR exerted cell proliferation, glycogen release, and antiinflammatory effect, confirmed by the decrease of IL-6 and the increase of both IL-10 and SIRT-1 expression levels. In conclusion, the present study provides new insight into developing a synbiotic formulation that could represent a promising supplement with health-promoting benefits suitable to support women's well-being.

The increased consumption of ready-to-eat fruits highlights the need for better control of microbial growth during their shelf life. Among bacteria, Staphylococcus aureus and Bacillus cereus are proposed as target species for testing alternative preservative methods. This study aimed to evaluate the antimicrobial effect of the cell-free supernatant (CFS) from LAB strains previously isolated from ready-to-eat fruits, used as a mixed solution, against both reference and native S. aureus and B. cereus, which were isolated from commercial ready-to-eat fruits. A specific challenge test was conducted on minimally processed orange slices, assessing the effect of CFS on the intentionally inoculated target bacteria using a culturing and quantitative PCR (qPCR) approach. Microbiological counts varied widely among samples, indicating an initial microbiota below legislative limits, mainly comprising total mesophilic and psychrophilic bacteria, which increased significantly after 8 days of storage. Additionally, our results demonstrated the food matrix’s capacity to support the growth of both target species, with the tested CFS mainly effective in reducing the growth of reference strains. The results of the physicochemical analyses showed that during refrigerated storage, the orange slices underwent changes in pH, color, and texture, mostly in S. aureus strain-inoculated samples, negatively affecting texture at mid-storage time. The study also underscored the importance of combining plate counting with qPCR methods to detect B. cereus, as it can be risky even at low levels.