Search Results (1,197)

The chemical composition of donkey and sheep milk showed significant differences, with donkey milk containing less fat and protein and a higher level of lactose, while sheep milk had more dry matter and a higher proportion of casein and saturated, unsaturated, monounsaturated, and polyunsaturated fatty acids. This study investigated the biological effects of in vitro-digested donkey and sheep milk, and their blends at different ratios, on differentiated human intestinal Caco-2 cells. Gene expression analysis showed that treatment with digested donkey milk (100%) significantly altered the expression of the autophagy-related gene (SQSTM1), tight junction (CLDN4), and innate defense genes (DEFB1, MUC2, and MUC5), showing a non-cytotoxic response and a profile consistent with a barrier-related transcriptional response. In contrast, sheep milk (100%) and certain milk blends (70% and 50% sheep’s milk) down-regulated genes involved in the autophagy process (ULK1, AMBRA, BECN1, ATG5, GABARAP, and SQSTM1), tight junction genes (OCLN and CDH1), and innate defense genes (DEFB1 and MUC2), suggesting a distinct epithelial response that warrants further confirmation. Metabolomic profiling revealed clear compositional and functional distinctions among the digested milks, identifying 166 metabolites that were differentially regulated (136 compounds were down-regulated and 30 up-regulated). Donkey milk digestion yielded a metabolite profile enriched in polar peptides and compounds with reported antioxidant associations, whereas sheep milk digestion showed a different pattern dominated by hydrophobic peptides and lipid-related metabolites. These findings indicate that, under the tested in vitro conditions, donkey milk digestion is associated with gene expression and metabolomic patterns consistent with epithelial barrier support. Full article

This study aimed to investigate the effects of thermo-mechanical treatment and solid-state fermentation (SSF) on the structural, physicochemical, and techno-functional characteristics of wheat bran (WB), with a particular emphasis on specific isolated protein fractions, starch, and dietary fibre. WB was subjected to extrusion at different temperatures (90, 115, and 130 °C) and screw speeds (16 and 25 rpm), followed by a 24-h SSF with Liquorilactobacillus uvarum. Changes in chemical composition, hydration properties (water absorption, solubility, swelling capacity), temperature-dependent (30–80 °C) extract viscosity, as well as protein extraction yields and in vitro protein digestibility of isolated protein fractions were systematically evaluated. Extrusion reduced insoluble dietary fibre (IDF) content by 15.7–33.9%, while increasing soluble dietary fibre (SDF) content by 59.1–94.2%, and SSF with L. uvarum additionally increased the SDF fraction by 6.5–7.9%. Regarding technological properties, extrusion increased WB water solubility (WS) up to 14.6%, and 24-h fermentation further enhanced WS, reaching up to 16.3%. Extrusion increased the degree of starch gelatinisation up to 49.1%, whereas fermentation reduced it to 40.7% and decreased the WB extract viscosity by 18.3–24.6%. Furthermore, 24-h SSF enhanced the in vitro digestibility of globulin, gliadin, and glutenin fractions from initial values of 71.3–78.8%, 70.6–75.2%, and 70.2–73.0% to 82.0–85.6%, 83.3–88.6%, and 79.4–89.0%, respectively. The observed thermo-induced thickening and enhanced digestibility suggest that modified wheat bran has potential as a functional bio-ingredient for future application in novel food formulations. Full article

Toad poison (ChanSu), a traditional animal-derived medicine, has long been used in East Asian medical systems for the treatment of inflammatory conditions and tumor-related disorders. While bufadienolides have been extensively investigated as its major bioactive constituents, the contribution of peptide components to its antitumor effects remains largely unexplored. Here, we identified protein-derived candidate antiproliferative peptides from toad poison using an integrated proteomics and bioinformatics-guided strategy. Proteomic analysis identified 135 proteins, from which 2117 peptide sequences were generated via in silico digestion with trypsin and pepsin. Subsequent multi-step screening using PeptideRanker, AntiCP, iACP, and ACPred yielded twelve candidate peptides with predicted anticancer activity. Network pharmacology analysis suggested their potential involvement in cancer-related targets and pathways. ADMET (Absorption, Distribution, Metabolism, Excretion, and Toxicity) evaluation was subsequently used as a complementary assessment of drug-like and safety-related properties, further prioritizing four peptides for experimental validation, while molecular docking supported their interactions with key lung cancer-associated targets. In vitro assays demonstrated that three of the four prioritized peptides (WEAWN, NSQWG, and ACGVIGICQ) exhibited initial antiproliferative activity against human lung cancer A549 cells, though these findings should be interpreted with caution given the absence of a positive control in the MTT assay. This study provides preliminary evidence suggesting that protein-derived peptide candidates from toad poison may possess antiproliferative potential, representing an early systematic exploration of its previously unexplored peptidome as a source of candidate antiproliferative peptides warranting further pharmacological investigation. The integrated strategy presented here offers an efficient approach for the discovery of bioactive peptides from animal-derived traditional medicines. Full article

Germinated Andean lupin whole flour (GAL) is rich in protein, dietary fiber, essential amino acids, and bioactive compounds, representing a promising alternative for the development of sustainable plant-based foods. This study investigated the feasibility of partially replacing soy protein isolate (SPI) with GAL in high-moisture extruded meat analogues. A central composite design was applied to evaluate the effects of the GAL ratio (0:100–50:50) and feed moisture content (50–70%) on the technological properties of the extrudates. The Response Surface Methodology was used to model and optimize the process. The incorporation of GAL significantly affected the (p < 0.10) water solubility index (WSI), oil absorption capacity (OAC), cooking loss (CL), yellowness (b*), cohesiveness, and adhesiveness, generating predictive models with satisfactory goodness-of-fit (R² > 0.75). Increasing GAL levels increased the WSI from 6.21 to 18.79% and cooking loss from 1.01 to 3.84%, while reducing OAC from 239.42 to 166.57%, indicating substantial modifications in matrix organization and hydration behavior. Numerical optimization identified an optimal formulation containing 12% GAL, 88% SPI, and 65.5% feed moisture, with a desirability of 77.82%. Model validation showed relative deviations lower than 10% between predicted and experimental values. The optimized meat analogue exhibited high protein content (84.44%), favorable techno-functional properties, and improved nutritional quality, with higher levels of branched-chain amino acids (17.34 g·100 g⁻¹ protein), essential amino acids (33.32 g·100 g⁻¹ protein), and in vitro protein digestibility (90.1%) compared with the control formulation. Multivariate analyses confirmed that phenylalanine, histidine, methionine, and leucine were the main variables that discriminated between the protein sources and the extruded products. Overall, GAL demonstrated strong potential as a sustainable functional ingredient to produce nutritionally enhanced high-moisture meat analogues. Full article

This study evaluates the effects of plant part (leaves vs. stems), harvest season (spring vs. autumn), and irrigation on the chemical composition, in vitro ruminal fermentation kinetics, and digestibility of three Mediterranean leguminous shrubs: Acacia fimbriata, Medicago arborea and Anthyllis barba-jovis. Samples were collected under field conditions, analysed for chemical composition, and assessed for in vitro ruminal fermentation, including incubations with and without polyethylene glycol (PEG) to assess the biological activity of phenolic compounds. Crude protein and fibre fractions did not differ significantly among species; ash was highest (p < 0.05) in Ac. fimbriata. In vitro fermentation kinetics were similar among species; however, the response to PEG was higher (p < 0.05) for Ac. fimbriata than for M. arborea and An. barba-jovis. In vitro digestibility was superior (p < 0.05) in M. arborea and An. barba-jovis. Leaves contained higher ash and protein contents and less fibre than stems, resulting in greater gas production, fermentation rate, and digestibility (p < 0.05). Spring-harvested fodder showed higher digestibility than autumn samples. Irrigation had negligible effects, except for a marginal numerical but nonsignificant (p = 0.05) greater extent of degradation observed in browse prior to irrigation. Overall, M. arborea showed a better nutritional quality, whereas Ac. fimbriata was the species with the lowest nutritive value. Forage quality was strongly influenced by plant part and harvest season, with leaves and fodder in spring showing superior potential as a feedstuff for ruminants. Full article

The gelation ability of gelatin is useful for improving texture and stabilizing food structure, but excessive gelation may limit its application in liquid and high-moisture food systems. Therefore, this study compared the effects of Alcalase and pepsin hydrolysis on the gel-forming ability, viscosity, and functional properties of porcine skin gelatin. o-Phthaldialdehyde (OPA) analysis showed that the Alcalase hydrolysate had a higher free amino group content (0.77 meqv/g protein) than the pepsin hydrolysate (0.23 meqv/g protein), indicating more extensive peptide bond cleavage (p < 0.05). The control formed a stable gel structure after 60 min at 25 °C, whereas both hydrolysates showed reduced gel-forming ability. In particular, the Alcalase hydrolysate was found to maintain high fluidity, completely inhibiting gel formation. Furthermore, 17.5% (w/v) Alcalase hydrolysate solution showed no measurable viscosity. In contrast, 17.5% (w/v) pepsin hydrolysate solution exhibited a viscosity of 653.88 cP. In contrast to the Alcalase hydrolysate, the pepsin hydrolysate also showed the highest oil absorption capacity (8.81 g/g), emulsion stability index (854.09 min), and a slightly higher in vitro digestibility (29.36%) (p < 0.05). These results demonstrate that enzyme-specific hydrolysis can differentially modify the balance between gel suppression and techno-functional properties of porcine skin gelatin. Full article

This study evaluated the effect of solid-state fermentation (SSF) using Aspergillus awamori on the nutritional, functional, antioxidant, structural, and protein-digestibility properties of black gram (Vigna mungo) flour and its application in wheat-based composite flour. Black gram was fermented for 0–12 days, and the 6-day fermented sample was selected for flour fortification based on improved nutritional and functional responses. SSF reduced tannin and phytic acid contents while increasing total phenolic content from 2.69 to 3.37 mg GAE/g, total flavonoid content from 0.83 to 1.65 mg QE/g, and DPPH radical-scavenging activity from 2.13 to 4.09 mg AAE/g by day 6. Protein content increased from 24.14% to 27.11%, and in vitro protein digestibility improved from 70.86% to 82.89%. Total amino acid content also increased from 88.71 to 98.24 g/100 g protein by day 6, followed by a decline during prolonged fermentation. Mineral contents, including iron, zinc, and calcium, were also improved after fermentation. Incorporation of 6-day fermented black gram flour into wheat flour enhanced the protein, crude fiber, water-holding capacity, oil-holding capacity, and in vitro protein digestibility of the composite flour. Overall, SSF with A. awamori improved the nutritional and functional quality of black gram and demonstrated its potential as a protein-rich ingredient for fortified cereal–legume composite flour development. Full article

The transition toward sustainable and nutrient-dense food systems requires novel plant-based ingredients and multifunctional formulations. In this study, gluten-free pasta was developed from millet (Panicum miliaceum) and psyllium husk (Plantago ovata), enriched with duckweed (Lemna minor) flour at 0–20% inclusion levels, with and without egg addition. The study aimed to evaluate the combined effects of duckweed incorporation and processing (drying, cooking, and in vitro digestion) on nutritional quality, bioactive compound stability, and volatile aroma composition. Duckweed enrichment (5–10–12–50% based on millet flour) contributed to strengthening the protein content, total polyphenol levels, and antioxidant capacity across all formulations. Processing steps markedly influenced compound stability and bioaccessibility: thermal treatments reduced extractable antioxidant activity, whereas simulated gastrointestinal digestion enhanced the bioaccessibility of bioactive compounds. Technological properties were moderately affected, with increasing duckweed levels resulting in longer cooking times and higher water absorption capacity. GC–MS–olfactometry analysis revealed that duckweed flour is characterized by a complex volatile profile dominated by hydrocarbons, aldehydes, alcohols, and sulphur-containing compounds, contributing primarily green, fatty, and herbaceous odor notes. During pasta processing, a substantial shift in aroma composition was observed, with ketones and alcohols becoming dominant, while key duckweed-derived odor-active compounds (e.g., carotenoid degradation products and green leaf volatiles) were partially retained in the final products. Olfactometric evaluation confirmed that although processing reduces aroma intensity, duckweed-related volatile markers remain detectable at higher incorporation levels. Overall, millet–psyllium–duckweed pasta represents a promising gluten-free functional food system combining improved nutritional value with plant-based protein enrichment and antioxidant potential. However, aroma modification during processing remains a critical factor for sensory optimization and consumer acceptance. The results highlight the importance of integrating nutritional, technological, and GC–MS–olfactometry approaches in the development of next-generation sustainable cereal-based foods. Full article

Common carp (Cyprinus carpio) fry were fed diets containing defatted Hermetia illucens (HI) larvae meal at three inclusion levels to investigate its effects on the nutriphysiological status, focusing on protein and fat digestibility (in vitro and in vivo), growth performance parameters, diet utilization, somatic indices, and GIT histomorphology. The HI meal was included at 0%, 2%, 4%, and 6%. A total of 240 carp fry were randomly divided into four groups, six replicates each (10 fish/tank), and the growth trials lasted 60 days. The dietary inclusion of HI meal up to 6% in carp feed did not significantly affect growth performance, feed utilization, or homeostasis. In vitro protein digestibility exceeded 97% across all treatments with pepsin, indicating efficient hydrolysis. However, the highest crude protein digestibility coefficients were recorded in all treatment groups, which differed significantly from the control group (90.5%). Also, villus width and crypt depth did not differ significantly; however, other parameters, such as villus height and area, showed some significant variations across the treatment groups. No adverse effects were observed on liver condition, structure, or function, reinforcing the safety of HI up to 6% in carp diets. In conclusion, this study demonstrates that HI can be incorporated as a functional, health-promoting ingredient in the diets of common carp fry. Full article

The narrow-clawed crayfish (Pontastacus leptodactylus), a non-native species accidentally introduced into China’s Irtysh River Basin, has formed a wild population with aquaculture potential, but its natural diet and protein utilization remain poorly understood. This study investigated its feeding ecology and digestive responses to protein sources to support ecological assessment and feed formulation. Intestinal contents and muscle samples from wild crayfish were analyzed using eDNA metabarcoding and fatty acid signature analysis. In vitro digestibility of 10 protein ingredients was evaluated with crude digestive enzyme extracts, and a 4-week trial compared an all-plant-protein diet (PPD) with an all-animal-protein diet (APD). eDNA indicated an omnivorous diet dominated by planktonic taxa, especially Rotifera, and fatty acid biomarkers suggested contributions from diatoms, dinoflagellates, phytoplankton, zooplankton, and benthic organisms. Soybean meal and soy protein concentrate showed high dry matter digestibility, whereas fishmeal and krill meal had high crude protein digestibility and amino acid release. Compared with PPD, APD increased feed intake, apparent nutrient digestibility, hemolymph nutritional indicators, villus height, and intestinal trypsin and lipase activities; however, intestinal integrity was preserved in both groups, and most intestinal antioxidant indices did not differ significantly. These findings indicate that animal proteins enhance nutrient utilization in P. leptodactylus, while selected plant proteins may serve as partial alternatives. Full article

Rapeseed meal (RSM) is an important protein source in animal feed, but its high levels of anti-nutritional factors and crude fiber (CF) limit its utilization in pigs. Our study compared the morphological structure, nutrient composition, and anti-nutritional factors of RSM, extruded rapeseed meal (ERM), and extruded solid-state enzymatically hydrolyzed rapeseed meal (EERM), and evaluated their digestibility in growing pigs. The results showed that ERM and EERM increased surface porosity. Notably, ERM and EERM had higher trichloroacetic acid-soluble protein contents and lower fiber, glucosinolate (GL), and isothiocyanate (ITC) contents. Compared to RSM, the in vitro digestibility of crude protein (CP) was significantly lower in ERM (p < 0.05), whereas EERM exhibited significantly higher digestibility of CP than ERM (p < 0.05). Additionally, compared to RSM and ERM, the digestibility of dry matter (DM) and gross energy (GE) increased significantly in EERM (p < 0.05). For in vivo digestibility, compared to ERM, the digestibility of CP and GE increased significantly in EERM (p < 0.05). In conclusion, our study demonstrates that ERM and EERM can change the chemical composition of RSM, and improve the digestibility of nutrients in pig production. Full article

Tilapia (Oreochromis niloticus) scales represent an underutilized by-product with considerable potential as a source of bioactive peptides that can be released through enzymatic hydrolysis. This study evaluated the production of protein hydrolysates from demineralized tilapia scales using Alkaline Protease or Esperase 8.0 L^® (Alkaline Protease and Esperase 8.0 L® were kindly provided by Novozymes, Bagsværd, Denmark), and examined the impact of simulated gastrointestinal digestion (SGID) on their bioactivities. The highest degree of hydrolysis (DH) was obtained with Alkaline Protease (12.4%), generating peptide fractions predominantly below 1000 Da, with a major population around 888 Da. Both hydrolysates exhibited antioxidant activity, with the Alkaline Protease hydrolysate showing higher ferric reducing antioxidant power (FRAP) and Fe(II)-chelating activity. The hydrolysates also displayed significant angiotensin-converting enzyme (ACE) inhibitory activity (IC₅₀: 13–14 µg/mL), dipeptidyl peptidase IV (DPP-IV) inhibitory activity (IC₅₀: 0.66–0.69 mg/mL), and prolyl endopeptidase (PEP) inhibitory activity (IC₅₀: 0.63–0.72 mg/mL). The digests were non-cytotoxic at the concentrations tested (<10 mg/mL) in STC-1 enteroendocrine cells. Following SGID, increased ACE-inhibitory activity was observed, with IC₅₀ values as low as 4.6 µg/mL, whereas DPP-IV and PEP-inhibitory activities decreased. The intestinal digest of the Esperase hydrolysate also exhibited significant cellular antioxidant activity in the ROS assay. Overall, these results indicate that tilapia scale hydrolysates are a promising source of peptides associated with in vitro enzyme inhibitory and antioxidant activities. However, the specific bioactive peptides responsible for these effects were not identified. Therefore, further studies involving peptide characterization, bioavailability assessment, and in vivo validation are required to establish their physiological relevance and potential applications as functional ingredients. Full article

Marine rest raw materials are often undervalued or wasted despite their nutrient and bioactive composition. Calanus finmarchicus, harvested primarily for its omega-3-rich oil, yields a side-stream protein hydrolysate, C. finmarchicus hydrolysate (CFH), during commercial enzyme-assisted extraction. Although currently used as a feed ingredient, CFH contains low-molecular-weight peptides and free amino acids with potential for human health applications. This study evaluated the gastrointestinal stability of CFH and the impact of digestion on bioactivity using a static in vitro gastrointestinal digestion model. Fresh-frozen and freeze-dried C. finmarchicus were included to provide comparative data. Antioxidant capacity was measured by ferric reducing antioxidant power (FRAP) and oxygen radical absorbance capacity (ORAC) assays, and antidiabetic activity by dipeptidyl peptidase-IV (DPP-IV) and protein tyrosine phosphatase 1B (PTP1B) inhibition assays. The hydrolysate maintained its antioxidant capacity throughout digestion (at 165 min: FRAP: 27.5 ± 0.6 µmol TE/g dry weight (DW); ORAC: 411 ± 37 µmol TE/g DW). Digestion increased its DPP-IV inhibitory activity, with the inhibitory concentration (IC₅₀) decreased from 3.73 to 1.96 mg/mL (p ≥ 0.05). PTP1B inhibitors were nonselective and detected only at 0 and 30 min. These findings support our hypothesis that CFH may serve as a nutraceutical for humans and provide a rationale for subsequent in vivo studies. However, further identification of bioactive components and in vivo validation are warranted. Full article

Whole-plant soybean (WPS) is a high-protein forage resource, but its natural ensiling is often unsatisfactory due to low water-soluble carbohydrate content and high buffering capacity. This study investigated the effects of exogenous melatonin (ME) at 0 (CK), 5 (ME1), 10 (ME2), and 20 (ME3) mg/kg fresh matter on fermentation quality, chemical composition, in vitro rumen fermentation, and bacterial community structure of WPS silage. ME2 and ME3 had lower pH values and higher lactic acid contents than CK, with both treatments achieving pH values below 4.2. Crude protein concentration increased from 15.42% in CK to 19.96% in ME3, while neutral detergent fiber was lower in all ME treatments, and acid detergent fiber was lower in ME2 and ME3 than in CK. At 36 h, no overall treatment effect was detected for cumulative gas production, whereas in vitro dry matter digestibility differed only between ME2 and ME3. 16S rRNA gene sequencing revealed that ME altered the bacterial community, with community-weighted rrn copy number elevated in ME2 and ME3. Random forest analysis identified Enterococcus as the genus with the highest importance for treatment classification, and functional predictions indicated higher predicted abundances of amino acid biosynthesis pathways in ME treatment groups. These results indicate that ME has potential as an additive for improving WPS silage fermentation, but practical dosage recommendations require further validation through aerobic stability, animal performance, economic, and safety assessments. Full article

Encapsulation serves as a critical strategy for the preservation of sensitive bioactive compounds, ensuring their stability and functionality within complex food matrices. Cellulose nanocrystals (CNCs) upcycled from by-products are being favoured as wall materials because they offer a sustainable yet powerful solution for maintaining compound stability. This study evaluated the encapsulation of fish oil (FO) within a nanocomposite matrix of garlic skin-derived cellulose nanocrystals (GCNCs) and soy protein isolate (SPI). The synergistic effects of FO loading and GCNC:SPI ratios on the microcapsules’ structural, physicochemical, and digestive properties were investigated. Higher FO loading significantly reduced the moisture content of the resulting microcapsule powders while increasing bulk and tapped densities by minimising internal porosity. Microstructural analysis showed irregularly shaped agglomerates. Higher FO loading also increased surface oil retention and inter-particle adhesion of the microcapsule powders; however, elevated SPI levels effectively counteracted these effects. Colour analysis further revealed that higher FO loading reduced powder lightness (L*) and increased yellowness (b*), while greater GCNC content positively influenced redness (a*). The formulation containing 10% FO, 3% GCNCs, and 7% SPI was identified as the optimal treatment. This ratio achieved the highest encapsulation efficiency (65.77% ± 1.10) and demonstrated superior flowability, characterised by the lowest Carr’s Index (20.65% ± 0.29) and Hausner Ratio (1.23 ± 0.05). Additionally, it maintained oxidative stability, with TBARS values (2.42 ± 0.08 mg MDA/kg oil) remaining consistently below the established 3 mg MDA/kg threshold. Fourier Transform Infrared Spectroscopy confirmed the successful entrapment of FO within the GCNC–SPI matrix. According to the in vitro digestion assays, the wall material provided a durable barrier in acidic media because the gastric release (28.04–55.28%) was significantly lower than the intestinal release (64.38–77.62%). The predominant fatty acids identified in both encapsulated and unencapsulated products were myristic acid (saturated fatty acid), elaidic acid (monounsaturated fatty acid), and docosadienoic acid (polyunsaturated fatty acid). Superior nutritional quality index (NQI) values in the encapsulated samples underscore the effectiveness of the wall material in providing a critical defence against fatty acid degradation and preserving overall oil quality. These findings suggest that the GCNC/SPI binary system is a highly effective delivery vehicle for protecting sensitive polyunsaturated fatty acids in functional food applications. Full article