Enzymatic Digestion Research Articles (Page 1)

Aquaculture temperatures vary depending on the fish species, and probiotics used in aquaculture must remain effective under these conditions. Therefore, our study developed host-associated low-temperature probiotics (HALP) adapted to temperature conditions relevant to aquaculture. Three bacterial strains, Rahnella inusitata NBL2302 (RI), Pseudoalteromonas arctica NBL2303 (PA), and Lactiplantibacillus plantarum NBL2306 (LP), were isolated from the gastrointestinal tract of Korean rockfish (Sebastes schlegelii). Safety assessments, in vitro probiotic characterization (including antioxidant activity, acid/salt/bile tolerance, antimicrobial activity, and adhesion), and a feeding trial were conducted. All three strains were confirmed to be safe, exhibiting no hemolytic or cytotoxic activity. Among the three strains, LP exhibited the highest adhesion to intestinal epithelial cells and showed antimicrobial activity against fish pathogens. RI supplementation significantly enhanced innate immune markers such as serum total protein (TP), triglycerides (TG), and myeloperoxidase (MPO), along with the upregulation of immune-related genes (HSP70, IL-1β, TNF-α) (p < 0.05). PA supplementation resulted in the greatest weight gain and significantly improved specific growth rate (SGR), likely due to increased digestive enzyme activity. LP promoted immunostimulatory responses (elevated expression of MPO, TP, HSP70, and TNF-α) and pronounced changes in the beta diversity of the gut microbiota. Microbiome analysis revealed that LP-fed fish harbored higher abundances of beneficial genera such as Pseudomonas, Ralstonia, and Sphingomonas. Overall, each strain displayed unique characteristics and exerted distinct effects during feeding trials. These findings, which take host temperature preferences into account, underscore the potential of HALP in aquaculture and highlight the need for further research into optimized combination strategies.

In the field of regenerative therapy, the stromal vascular fraction (SVF) extracted from adipose tissue has been widely recognized for its significant benefits. However, the cellular composition and therapeutic effect of SVF products prepared via different methods are unclear. SVF cells were obtained via three approaches: (1) generation of the SVF via mechanical emulsification (M-SVF), (2) generation of the SVF via laboratory enzymatic digestion (L-SVF), and (3) generation of the SVF via commercial cell separation systems (C-SVF). We evaluated their healing effects on mouse wounds. Additionally, we utilized single-nucleus RNA sequencing (snRNA-seq) technology to explore the cellular composition of the C-SVF. The cell yield of C-SVF was comparable to that of L-SVF. During in vitro culture, C-SVF exhibited enhanced proliferation and a reduced proportion of apoptotic cells. In a mouse wound model, the application of C-SVF facilitated the closure of mouse wounds and improved collagen remodeling and angiogenesis in the wound area. Additional snRNA-seq analysis revealed that APOE + adipose-derived stem cells and immune cells, especially M2 anti-inflammatory macrophages, are enriched in C-SVF, which together promote wound repair, and that APOE + adipose-derived stem cells (ADSCs) and immune cells, especially M2 anti-inflammatory macrophages, are enriched in C-SVF, which jointly regulate and promote wound repair. A commercial extraction system is an effective method for isolating viable SVF cells enriched with APOE + ADSCs and M2 macrophages.

Abstract Lignocellulosic biomass (LCB), such as agricultural residue rich in cellulose and hemicellulose, can serve as a feedstock for bioethanol production due to its environmental benefits. Deep eutectic solvents (DES) are low-cost and greener solvents for LCB pretreatment. The present study investigated the efficiency of two DES types for rice straw pretreatment under microwave conditions: choline chloride:glycerol (ChCl:Gly) and choline chloride:formic acid (ChCl:FA). DES pretreatment was performed under microwave conditions (100–140 °C and 5–15 min) at 10% solids loading and constant power 200W. After pretreatment, the samples were characterized, and enzymatic digestibility was investigated at 50 °C for 72 h using enzyme loading of 6 filter paper unit g −1 cellulose and 10% solids loading. Untreated rice straw consists of cellulose (41.8%), hemicellulose (24.9%), lignin (17.0%) and ash (15.0%). The cellulose content increased to 59.8% and 59.2% after ChCl:Gly and ChCl:FA pretreatment, respectively, while hemicellulose decreased to 15.6% and 10.1% after ChCl:Gly and ChCl:FA pretreatment, respectively. Lignin content decreased to 8.2% after ChCl:Gly pretreatment compared to 9.6% after ChCl:FA pretreatment. Ash content (20.4%) obtained after ChCl:FA pretreatment was higher than 14.0% obtained after ChCl:Gly pretreatment. The cellulosic and hemicellulose fractions from ChCl:Gly pretreatment were effectively hydrolyzed with glucose and xylose yield of 86.0% and 68.4%, respectively, compared to glucose and xylose yield of 58.2% and 62.8%, after ChCl:FA pretreatment. ChCl:Gly pretreatment enriched cellulosic content and preserved hemicellulose fraction, achieving a higher yield of fermentable sugars than ChCl:FA pretreatment; hence, it can potentially support a biorefinery.

The growing global demand for clean energy and sustainability has increased interest in lignocellulosic biomass as a viable alternative to conventional fossil fuels. Among the various biomass resources, sugarcane bagasse, an abundant agro-industrial by-product, has emerged as a promising feedstock to produce renewable fuels and value-added chemicals. Its high carbohydrate content offers significant potential for bioconversion. However, its complex and recalcitrant lignocellulosic matrix presents significant challenges that necessitate advanced pretreatment techniques to improve enzymatic digestibility and fermentation efficiency. This review consolidates recent developments in the valorization of sugarcane bagasse focusing on innovative pretreatment and fermentation strategies for sustainable bioethanol production. It emphasizes the synergistic benefits of integrating various pretreatment and fermentation methods to improve bioethanol yields, reduce processing costs and enhance overall process sustainability. This review further explores recent technological advancements, the impact of fermentation inhibitor, and emerging strategies to overcome these challenges through microbial strains and innovative fermentation methods. Additionally, it highlights the multi-faceted advantages of bagasse valorization, including waste minimization, renewable energy production and the promotion of sustainable agricultural practices. By evaluating the current state of research and outlining future perspectives, this paper serves as a comprehensive guide to advancing the valorization of sugarcane bagasse in the transition towards a low-carbon economy. The novelty of this review lies in its holistic integration of technological, economic, and policy perspectives, uniquely addressing the scalability of integrated pretreatment and fermentation processes for sugarcane bagasse, and outlining practical pathways for their translation from laboratory to sustainable industrial biorefineries within the circular bioeconomy framework.

Abstract Purpose Echinoderms, such as the common sea star ( Asterias rubens , Asteroidea), are a disregarded and underestimated source of high protein biomass. They form locally large densities, cause damage to natural mussel beds and shellfish aquaculture, and constitute a large proportion of bycatch in fisheries. Currently, captured or fished sea stars are dumped or used as fertilisers. This study investigates the valorisation of sea star meal (SM) as an alternative to fishmeal (FM) in aquaculture diets for Whiteleg shrimp ( Litopenaeus vannamei ). Methods Common sea stars were processed to meal and incorporated into four experimental diets with increasing levels of SM, replacing 0, 20, 50, and 80% of FM. The diets were fed to shrimp for 42 days in a recirculating aquaculture system. Survival, feed intake, growth, hepatosomatic index, haemolymph metabolic, and immune parameters as well as apparent digestibility and digestive enzyme activities were determined (Animal trial Vet Authority registration number/date: TA21-01/31.08.2021). Results All treatments showed equally high survival rates and feed intake. Shrimp showed maximum growth on the diet with the highest SM inclusion (80%), which significantly exceeded growth of shrimp in the 0% replacement diet. The feed conversion ratio was significantly lower for shrimp fed the 50% and 80% SM-diets than the 0%-diet. Protein and energy digestibility were significantly higher in the 80% SM-diet compared to the other diets. Metabolic and immune parameters were not affected by the different diets. Conclusion Sea star, A. rubens , biomass shows high potential as a “sustainable added-value material” for shrimp feed. Its use can reduce the dietary FM content to 5.4%. Therefore, the authors recommend application in novel value chains for aquaculture in Europe. Trial Registration The animal experiment was approved and registered by the local veterinary authority in Bremen, Germany (approval ID: TA21-01, date of approval: 31.08.2021). Graphical Abstract

A forgotten traditional superfood is the best breakfast on those days has always played an important role in everyone’s life duringolden ancestor period it was never about eating fancy food but something which is wholesome and heathywhich is our energy booster watery diet (pazhayasooru kanji)-NEERAGARAM. Researchers have conducted many studies on the diet states it gives more energy booster starting from adulthood to geriatric. The fermentation process helps the probiotic ,lactic acid bacteria are known to improve gut health, increase the bioavailability of electrolytes, aid in digestion,produce B-vitamins and beneficial enzymes and also it gives special attention to the diabetes patient where the blood sugar regulates when drinking this super drink.“Fermented rice is called “Pazhayasaadam” in Tamil, “Vellachoru” in Malayalam, and “Chaddannam” in Telugu. The rice water is called “Neeragaram” in Tamil.”

This study was conducted in two phases: first, to assess the impact of microbial fermentation on enhancing the nutritional quality of fish by-products, and second, to evaluate the effects of replacing fishmeal with these fermented by-products in the diet of Nile tilapia (Oreochromis niloticus) on growth performance, blood parameters, antioxidant indices, immunity, digestive enzyme activity, and carcass composition. In the initial phase, proteolytic activity of five bacterial strains including Bacillus subtilis (ATCC: 6051), B. licheniformis (IBRCM: 10204), Lactiplantibacillus plantarum (PTCCs: 1058 and 1745), and Lactobacillus casei (PTCC: 1608) was evaluated using growth assays in skimmed milk culture media and analyzed using Image-J software. B. licheniformis exhibited the highest proteolytic activity and was selected for fermentation. Resulting hydrolyzed proteins were characterized by peptides with molecular weights below 11 kDa. In the second phase, fishmeal was replaced with fermented by-products at five levels (0 (control), 25, 50, 75, and 100%). Two hundred ten Nile tilapia with an average weight of 2.83 ± 0.05 g were stocked in fifteen 200 L plastic tanks at three replicates, with 14 fish per tank, and fed daily at a rate of 7% of their body weight for 63 days. With increasing levels of fishmeal replacement (25% to 75%), significant improvements (p &lt; 0.05) were observed in final weight gain, body weight gain, specific growth rate, protein production value, and protein efficiency ratio. Additionally, blood plasma concentrations of hormones T3 and T4, immunoglobulin level, the activities of complement (ACH50), and antioxidant enzymes (catalase and superoxide dismutase) increased significantly in fish fed the diets with fermented by-products compared to those of the control diet (p &lt; 0.05). The optimal replacement levels for specific growth rate and feed conversion ratio were identified as 86.28% and 83.91%, respectively.

This study investigated the effects of dietary supplementation with composite acidifying agents containing 2-hydroxy-4-methylthiobutyric acid (≥30.0%), lactic acid (≥24.2%), and phosphoric acid (≥23.8%) on production performance, egg quality, serum biochemistry, intestinal health, and cecal microbiota in 300-day-old BIAN chickens. In a 42-day randomized trial, 900 laying hens were randomly allocated to three groups: the control group (basal diet with tap water), test group A (basal diet with 0.05% composite acidifier in drinking water), and test group B (basal diet with 0.20% composite acidifier in drinking water). The results demonstrated that test group B exhibited a significant 4.6% increase in average egg weight compared to the control (p = 0.029), while test group A showed enhanced Haugh unit values (p = 0.010) and eggshell strength (p = 0.010). Serum biochemical analysis revealed marked improvements in immune function, with test group B showing a 65.49% increase in globulin levels (p = 0.010) and 61.76% elevation in total antioxidant capacity (T-AOC) (p = 0.010). Intestinal digestive enzyme activities were significantly enhanced, particularly in test group A with a 61.73% increase in duodenal lipase activity (p = 0.010) and 37.43% elevation in jejunal amylase activity (p = 0.036). Morphological assessment demonstrated improved intestinal architecture in test group B, with a 26.02% reduction in crypt depth (p = 0.025) and a 44.53% increase in the villus-to-crypt ratio (p = 0.030). Microbiota analysis revealed dose-dependent modulation of cecal bacterial communities, with notable increases in beneficial genera including Akkermansia (from 1.8% to 7.2% in test group A) and Lachnospiraceae (from 4.7% to 9.7% in test group B) while maintaining core microbiota stability. Principal component analysis confirmed distinct microbial ecological niches created by acidifier supplementation. These findings demonstrate that composite acidifying agents effectively enhance egg production quality, immune status, digestive function, and gut health in BIAN chickens, supporting their potential as sustainable alternatives to antibiotic growth promoters in laying hen production systems.

Low-fishmeal feed is increasingly being adopted across the global aquaculture industry. This study evaluated dietary Clostridium butyricum and alanyl-glutamine (Ala-Gln) supplementation in juvenile triploid rainbow trout (Oncorhynchus mykiss) with a low-fishmeal diet. Four diets were tested: basal diet (SBM, 15% fishmeal and 21.6% soybean meal), SBM + 0.5% C. butyricum (CB), SBM + 1.0% Ala-Gln, and SBM + 0.5% C. butyricum + 1.0% Ala-Gln (CB-AG). Fish were fed in 500 L tanks in recirculating aquaculture systems for 8 weeks (62.52 ± 0.47 g). Each group comprised three tanks, with each tank housing 30 fish. Then 10 fish per tank were challenged with Aeromonas salmonicida. CB-AG showed significantly higher weight gain and specific growth rates than the SBM group (p &lt; 0.05). Mortality was significantly lower in CB-AG and AG than in SBM after A. salmonicida challenge. Histomorphology revealed significant differences (p &lt; 0.05) between CB-AG and SBM in muscularis thickness, villus width, and height. SBM sections showed inflammatory infiltration and border damage were attenuated in supplemented groups. Serum malondialdehyde (MDA) and dioxygenase (DAO) were significantly lower in CB-AG than SBM (p &lt; 0.05), while serum and hepatic lysozyme (LZM) and hepatic superoxide dismutase (SOD) were higher. Digestive enzymes indicated significantly higher trypsin and lipase activities in CB-AG (p &lt; 0.05). CB-AG upregulated intestinal tight junction proteins and PepT1 and downregulated pro-inflammatory mediators. Combined 0.5% C. butyricum and 1.0% Ala-Gln inclusion effectively preserved growth performance, antioxidant capacity, gut microbiome homeostasis, and intestinal health in rainbow trout on low-fishmeal diets.

Propolis, a natural resinous compound with bioactive properties, has gained attention as a potential growth stimulant in aquaculture. This study investigated the effects of dietary propolis supplementation on growth performance, molecular biomarkers, intestinal histomorphometry, and digestive enzyme activity in tilapia juveniles (Oreochromis niloticus), and saved 50% of the amount of fish meal used in feed. A total of 210 fish (15.55 ± 0.21 g) were distributed into 21 aquaria and fed six isonitrogenous (30% crude protein) and isocaloric (1820 kJ/kg) diets containing graded propolis levels (0, 2, 4, 6, 8 and 10 g/kg) for 86 days. We added two controls for the experiment, the first to assess propolis efficiency with and without its supplementation, and the second to monitor its plant-based protein substitution ability of 50% fish meal. Results indicated that 0.4% and 0.6% propolis supplementation significantly enhanced growth metrics, including final body weight, weight gain, specific growth rate, and feed conversion ratio, comparable to the control (B) diet (p < 0.05). Propolis inclusion also improved protease and amylase activities, with the highest enzymatic activity observed at 0.4% supplementation. Molecular analysis revealed upregulation of hepatic IGF-1 gene expression in the 0.6% propolis group, suggesting enhanced growth potential. Intestinal histomorphometry demonstrated increased villus height, width, and surface area in propolis-fed fish, indicating improved nutrient absorption. Furthermore, whole-body proximate analysis showed optimised protein deposition in 0.4% and 0.6% treated groups. These findings suggest that propolis at 0.4%-0.6% dietary inclusion can increase the effectiveness of plant protein mixture to replace half amount of dietary fishmeal, enhance digestive efficiency, and stimulate growth in tilapia juveniles, offering a sustainable alternative for aquaculture feed optimisation.

In-capillary enzymatic digestion beyond trypsin for the sensitive targeted bottom-up analysis of protein biomarkers by capillary electrophoresis-mass spectrometry.

Microplastic contamination in sediments: Analytical techniques and case-based evaluations.

In situ generation of peracetic acid using phosphotungstic acid: A one-pot strategy for selective delignification of corn stover.

Microplastic residue in recycled food co-products from mechanical depacking systems: a simulation study for animal food.

Phenolic composition, antioxidant capacity and digestive enzymes inhibition of Butiá (Butia catarinensis, Butia eriospatha, and Butia odorata) under simulated digestion.

Allergenic mites excrete cathepsins B and C as active cysteine proteases distinct from group 1 allergens.

Ziziphus spina-christi has been a part of traditional medicine for centuries in North Africa and the Middle East. Numerous secondary metabolites have been isolated from this plant, such as alkaloids, flavonoids, polyphenols, vitamin C and bioactive polysaccharides. Therefore, using Z. spina-christi leaf extract (ZSLE) is one of the recommendations for improving poultry performance and health. The goal of this study was to explore the influences of ZSLE as a feed additive on broiler growth, carcass traits, digestive enzymes, immunological and antioxidant parameters, lipid profile and blood metabolites. A total of 250 one-week-old Arbor Acres broilers were distributed into 5 groups; each group contained 5 replicates with 10 birds each. The treatments were (1) basal diet (no additive, ZSLE-0), (2) basal diet+2g ZSLE/kg (ZSLE-2), (3) basal diet+4g ZSLE/kg (ZSLE-4), (4) basal diet+6g ZSLE/kg (ZSLE-6) and (5) basal diet+8g ZSLE/kg diet (ZSLE-8). Z. spina-christi reduced feed intake while improving body weight gain and feed conversion ratio (FCR) (p<0.05). During the 3-6 and 1-6 weeks, birds fed ZSLE-4 improved FCR compared with the other birds (p<0.05). Compared to ZSLE-0, ZSLE-6 and ZSLE-8 had substantially greater gizzard and giblet percentages. However, ZSLE had no effect on carcass, dressing, liver or heart percentages. Birds fed ZSLE-4, ZSLE-6 and ZSLE-8 showed increases in amylase, lipase, protease, haemoglobin % (Hb), red blood cells (RBCs) and white blood cells (WBCs) count (p<0.05). ZSLE supplementation had no effect on aspartate aminotransferase (AST) (p>0.05), whereas ZSLE-4 and ZSLE-6 decreased alanine aminotransferase (ALT) levels (p<0.05). All ZSLE-supplemented groups showed improvements in total protein, globulin and lipid profiles. ZSLE significantly decreased cholesterol, low-density lipoprotein (LDL) and LDL/high-density lipoprotein (HDL) ratio. HDL was increased in ZSLE-4, ZSLE-6 and ZSLE-8 (p<0.05). Birds fed ZSLE-6 and ZSLE-8 improved the immunoglobulins (IgA, IgY and IgM), bursa of Fabricius%, thymus% and spleen% compared with the other birds (p<0.05). Additionally, lysozyme and C3 showed a significant linear influence. All ZSLE-supplemented groups showed low blood glucose, creatinine, uric acid and malondialdehyde (MDA) (p<0.05). ZSLE improved antioxidant capacity, and ZSLE-6 and ZSLE-8 showed the highest reduced glutathione (GSH), catalase (CAT) and total antioxidant capacity (TAC) levels (p<0.05). Collectively, ZSLE exhibits growth-promoting, immunostimulant, antioxidant and hypolipidemic properties and can be employed as a natural extract in the diet of broiler chickens.

Probiotic encapsulation strategies for controlled intestinal delivery and microbiome dysbiosis therapy.

Microbial dysbiosis and hepatic inflammation under combined pesticide exposure: Insights into gut-liver axis disruption in Japanese medaka.

Functional reorganization and systemic toxicity induced by microplastics and climate-relevant stressors in Tribolium castaneum: Are we simulating tomorrow's toxic reality?