Manuela Amorim

Background: Growing public health concerns regarding the consumption of conventional dairy products, coupled with the increasing prevalence of plant-based and vegan dietary preferences, have stimulated considerable scientific interest in the development of innovative fermented legume-based beverages. The lima bean (Phaseolus lunatus) represents a particularly promising raw material in this context, due to its high protein content, the presence of prebiotic α-galactosides, and its notable endogenous antioxidant compounds. Aims: This study aimed to evaluate the probiotic viability, physicochemical and biochemical properties, sensory characteristics, and antioxidant potential of fermented lima bean-based beverages subjected to refrigerated storage at 4°C for 28 days. Material and Methods: Four lima bean-based beverages (Lbs) were formulated by incorporating a 1% inoculum of either the single-strain probiotic Lactobacillus acidophilus La5 (LbLa5), Bifidobacterium animalis subsp. lactis Bb12 (LbBb12), a co-culture of both strains (Lb coculture), or no bacterial inoculum, serving as a non-fermented control beverage. Results: The probiotic Lbs exhibited a high crude protein content (3.15% – 3.34%) comparable to that of soy-based and bovine milk, and were characterized by a substantial carbohydrate fraction (2.08% – 2.32%). Viable counts of L. acidophilus La5 and B. lactis Bb12 were maintained at 6 – 7 log CFU/mL throughout the shelf-life period, with B. lactis Bb12 demonstrating enhanced survival under co-culture conditions. High-performance liquid chromatography (HPLC) analysis revealed significant reduction in the prebiotic oligosaccharide raffinose throughout storage with final concentrations ranging from 2.17 to 3.35 g/L. Probiotic metabolic activity induced a progressive decline in pH (4.00 – 4.30) accompanied by an increase in titratable acidity (0.70 – 0.89%). Chromatographic profiles demonstrated that both selected strains were proteolytically active during cold storage, resulting in a reduction of free amino acid content and the generation of bioactive low-molecular-weight peptides (≤ 10 kDa). Fermentation promoted an increase in total phenolic compound (TPC) content over the first seven days, whilst DPPH radical scavenging activity increased over the initial three weeks of storage. The co-culture treatment yielded the highest TPC content (1.71 – 2.03 mg GAE/mL) and DPPH radical scavenging activity (0.86 – 1.52 mg TE/mL) throughout the entire storage period. Sensory flavor attributes and overall acceptability were enhanced across all fermented beverage formulations relative to the non-fermented control. Conclusions: The incorporation of L. acidophilus La5 and B. lactis Bb12 into lima bean-based beverages yields a nutritionally dense, functionally enriched, and sensorially acceptable plant-based alternative well-suited to vegan and health-conscious dietary regimens. These findings support the valorization of Phaseolus lunatus as a viable substrate for the development of probiotic fermented beverages. Keywords: Lima Bean-Based Beverage; Probiotics; Raffinose; Organic Acids; Free Amino Acids; Bioactive Peptides; Antioxidant Capacity.

This study chemically characterized three Pleurotus ostreatus fruiting bodies cultivated in the Iberian Peninsula under different conditions (biological and industrial), with emphasis on polysaccharide analysis. Comprehensive comparative data on cultivation-dependent nutritional variations will potentially improve their nutritional and therapeutic applications. Industrial mushrooms (POC and POA) contained significantly higher carbohydrate content (74%), while the biologically cultivated mushroom (POL) exhibited more protein (22.6%), fat (4.2%), and ashes (8.0%). Monosaccharide analysis showed glucose dominance (28.7–45.5%), with mannose, galactose, xylose, and arabinose also present. Trehalose was the primary free sugar (4.8–14.9%). The (1→3)(1→6)-β-glucans varied significantly across samples (POL: 20.5%; POC: 29.3%; POA: 34.3%). Nuclear magnetic resonance analysis suggested complex polysaccharide arrangements. Water-soluble carbohydrates and proteins showed molecular weight distributions of 0.18–21 kDa and 0.20–75 kDa, respectively. All mushrooms were rich in essential amino acids, phosphorus (2.79–3.07%), potassium (0.56–0.68%), linoleic acid (0.82–1.14%), and oleic acid (0.22–0.31%). Fourier transform infrared confirmed a mushroom-specific biochemical profile. These findings corroborate the high nutritional value of POL, POC, and POA, with a significant contribution to the daily requirements of fiber, protein, and minerals (phosphorus, potassium, magnesium, iron, zinc, copper, and selenium), making them suitable for functional foods and nutraceuticals with cultivation-dependent nutritional profiles.

Cannabinoids have long been known for their bioactive properties, with their topical application as anti-inflammatory compounds being at the forefront of research for the past decade. Concurrently, the cosmetic market is a fast-growing industry in constant need of new biomolecules. In this work, we studied the safety profile for topical applications of two cannabinoids: cannabidiol (CBD) and cannabigerol (CBG) and assessed their potential as skin care ingredients. The CBG used in this work resulted from bio-fermentation, and to the best of our knowledge, there are no extensive reports on its safety and usage as a cosmetic ingredient. Our results show that CBD and CBG do not exhibit cytotoxicity, mutagenicity, or skin sensitization. Moreover, we verified an absence of primary irritability, accumulated irritability, phototoxicity and photosensitization, supporting the claims of dermatologically tested, hypoallergenic and non-irritating. While these cannabinoids did not show significant anti-aging effects by altering the extracellular matrix components (both in vitro and ex vivo), they demonstrated promise as protective agents against inflammation caused by air pollution. Specifically, they reduced the levels of pro-inflammatory cytokines, making them valuable in combating environmental skin damage. Overall, our results validate the safety of topical use of cannabinoids, while paving the way for further research in the beauty and personal care market as soothing agents.

Annona muricata is a fruit species belonging to the Annonaceae family, which is native to the warmer tropical areas of North and South America. A large amount of discarded residue from A. muricata is of interest for obtaining new industrial inputs. To propose the applications of the biopolymer from A. muricata residues (Biop_AmRs), this study aimed to characterize this input chemically and functionally, as well as to evaluate its potential for hemocompatibility and cytotoxicity activity in vitro. Biop_AmRs is an anionic heteropolysaccharide composed of glucose, arabinose, xylose, galactose, mannose, uronic acid, and proteins. This biopolymer exhibited a semicrystalline structure and good thermal stability. Biop_AmRs exhibited excellent water holding capacity, emulsifying properties, and mucoadhesiviness and demonstrated hemocompatibility and cytocompatibility on the L929 cell line. These results indicate possible applications for this biopolymer as a potential environmentally friendly raw material in the food, pharmaceutical, biomedical, and cosmetic industries.

Cannabidiol (CBD) and cannabigerol (CBG) are two pharmacologically active phytocannabinoids of Cannabis sativa L. Their antimicrobial activity needs further elucidation, particularly for CBG, as reports on this cannabinoid are scarce. We investigated CBD and CBG’s antimicrobial potential, including their ability to inhibit the formation and cause the removal of biofilms. Our results demonstrate that both molecules present activity against planktonic bacteria and biofilms, with both cannabinoids removing mature biofilms at concentrations below the determined minimum inhibitory concentrations. We report for the first time minimum inhibitory and lethal concentrations for Pseudomonas aeruginosa and Escherichia coli (ranging from 400 to 3180 µM), as well as the ability of cannabinoids to inhibit Staphylococci adhesion to keratinocytes, with CBG demonstrating higher activity than CBD. The value of these molecules as preservative ingredients for cosmetics was also assayed, with CBG meeting the USP 51 challenge test criteria for antimicrobial effectiveness. Further, the exact formulation showed no negative impact on skin microbiota. Our results suggest that phytocannabinoids can be promising topical antimicrobial agents when searching for novel therapeutic candidates for different skin conditions. Additional research is needed to clarify phytocannabinoids’ mechanisms of action, aiming to develop practical applications in dermatological use.

Pineapple by-products (peels and stems) from fruit processing industries were evaluated to understand its potential application as a functional food. Therefore, the bioactive compounds of pineapple by-products were characterized for prebiotic and antioxidant activities. A total characterization of soluble carbohydrates profile (simples and complex carbohydrates), as well as polyphenols was performed, after removal of enzymatic fraction from pineapple crude juice, allowing the decrease of proteolytic activity and improving the other biological activities. Results showed that pineapple liquid fraction, from stem and peels, can be applied as a prebiotic enhancer, promoting the growth of five probiotic microorganisms (two strains of Lactobacillus sp. and three strains of Bifidobacterium sp.), as a single carbohydrate source. Moreover, through HPLC (High Performance Liquid Chromatography) analysis, 10 polyphenols were identified in pineapple liquid fractions, with some expected differences between both evaluated by-products. Gastrointestinal tract was simulated, in a continuous mode to understand the impact of pH changes and gastrointestinal enzymes into pineapple liquid fractions. Results showed a digestion of high molecular weight polysaccharides into small molecular weight tri-, di-, and monosaccharides. There was an increase of samples antioxidant activity through the gastrointestinal stage, followed by the release of specific polyphenols, such as chlorogenic, coumaric, and ferulic acids. The prebiotic activity did not improve throughout the simulation, in fact, the prebiotic potential decreased throughout the different stages.