Protein Fractions Research Articles

Defining physicochemical properties of urinary proteins that determine their inhibitory activities against calcium oxalate kidney stone formation

We have recently reported a set of urinary proteins that inhibited calcium oxalate (CaOx) stone development. However, physicochemical properties that determine their inhibitory activities remained unknown. Herein, human urinary proteins were chromatographically fractionated into 15 fractions and subjected to various CaOx crystal assays and identification by nanoLC-ESI-Qq-TOF MS/MS. Their physicochemical properties and crystal inhibitory activities were subjected to Pearson correlation analysis. The data showed that almost all urinary protein fractions had crystal inhibitory activities. Up to 128 proteins were identified from each fraction. Crystallization inhibitory activity correlated with percentages of Ca2+-binding proteins, stable proteins, polar amino acids, alpha helix, beta turn, and random coil, but inversely correlated with number of Ox2−-binding motifs/protein and percentage of unstable proteins. Crystal aggregation inhibitory activity correlated with percentage of stable proteins but inversely correlated with percentage of unstable proteins. Crystal adhesion inhibitory activity correlated with percentage of stable proteins and GRAVY, but inversely correlated with pI, instability index and percentages of unstable proteins and positively charged amino acids. However, there was no correlation between crystal growth inhibitory activity and any physicochemical properties. In summary, some physicochemical properties of urinary proteins can determine and may be able to predict their CaOx stone inhibitory activities.

Bioactive and Nutritional Potential of an Infant Food Based on Mangifera Indica, Musa Paradisiaca, Chenopodium Quinoa and Amaranthus Caudatus Flour

There are few studies in the scientific literature on Peruvian native cereal-based instant mixes. Food aid programs of the Peruvian government allow food to be distributed to infants in public schools, and in general, these foods contain carbohydrates and a protein fraction, but there is some rejection on the part of consumers due to the sensory aspect of the product that does not has been considered in this population. The objective of this study is to know the bioactive, nutritional potential and the level of sensory acceptance of infant food made from Andean pseudocereals from the Ancash region and tropical fruits from the Piura region, Peru. Three formulations were prepared based on mango, banana, quinoa and kiwicha flours (T1, T2 and T3) and were compared with a commercial product based on kiwicha and oatmeal. The total amounts of phenolic compounds, vitamin C, antioxidant activity, composition, water absorption index, oil, milk was determined; viscosity and sensory evaluations were carried out with the participation of preschool children (4 and 5 years). The content of phenols, ascorbic acid (AA) and antioxidant capacity of T1 and T2 were 10 times higher than the control treatment. Physical and chemical composition analysis showed that there were no significant differences among the treatments evaluated. Banana flour contributes to the water and oil absorption capacity and viscosity. Finally, the addition of quinoa and kiwicha flour improves milk absorption capacity. The characteristics of the developed mixtures allow the incorporation of more food and less water to increase viscosity acceptance. The greatest preferences were obtained by T2 and T1, obtaining a level of acceptance above 82%, which shows that today children have a healthy food alternative such as mixtures of Andean pseudocereals and tropical fruits without sacrificing their nutritional qualities.

A Carrier Phase Ultrafiltration and Backflow Recovery Technique for Purification of Biological Macromolecules.

A simple carrier phase based ultrafiltration technique that is akin to liquid chromatography and is suitable for medium-to-large volume sample preparation in the laboratory is discussed in this paper. A membrane module was integrated with a liquid chromatography system in a "plug and play" mode for ease of sample handling, and recovery of species retained by the membrane. The sample injector and pump were used for feed injection and for driving ultrafiltration, while the sensors and detectors were used for real-time monitoring of the separation process. The concentration of retained species was enriched by utilizing controlled concentration polarization. The recovery of the retained and enriched species was enhanced by backflow of carrier phase through the membrane using appropriate combination of valves. The backflow of carrier phase also cleaned the membrane and limited the extent of membrane fouling. Proof-of-concept of the proposed technique was provided by conducting different types of protein ultrafiltration experiments. The technique was shown to be suitable for carrying out protein fractionation, desalting, buffer exchange and concentration enrichment. Adoption of this approach is likely to make ultrafiltration easier to use for non-specialized users in biological research laboratories. Other advantages include enhanced product recovery, significant reduction in the number of diavolumes of buffer needed for conducting desalting and buffer exchange, minimal membrane fouling and the potential for repeated use of the same module for multiple separation cycles.

Multitechnique approaches for characterising phase behaviour in food biopolymer systems

SummaryUnderstanding the phase behaviour of biopolymer composites is vital for various technological applications, such as developing functional foods, nutraceuticals, encapsulation, isolation and fractionation of enzymes and proteins. The growing consumer demand for healthier food products pressures the food and nutraceutical industries to innovate while maintaining traditional texture and nutritional quality. This review critically examines various analytical techniques, including spectroscopic, microscopic, calorimetric, rheological, viscometry, and diffraction methods, used to study phase behaviour. Each technique provides unique insights into phase behaviour mechanisms at micro and macroscopic levels, with respective advantages and limitations discussed. The review advocates for a multi‐technique approach for a comprehensive understanding of phase behaviour and highlights the development of in situ probing techniques with minimal sample pre‐processing as a promising advancement. These innovations are expected to significantly enhance the practical applications of these techniques in the food industry, meeting current consumer demands without compromising traditional qualities.

Substantiating the feasibility of using hemp seed protein in cooked sausage technology

The object of this study was the technology of cooked sausages with hemp seed protein. The research considers determining the effectiveness of using protein from hemp seeds in the technology of cooked sausages to increase the nutritional value and improve the functional and technological indicators when replacing part of the meat raw material. The subject of the research was meat model systems, cooked sausage with hemp seed protein. Three prototypes of cooked sausages based on the analog were developed. In experimental samples of cooked sausages, second-grade beef was replaced with protein from hemp seeds (Cannabis Sativa L.) (ToV "Desnaland", Ukraine) in amounts of 12, 14, and 16 %. The functional and technological indicators of hemp seed protein were previously investigated at hydromodules of 1:1, 1:2, and 1:3. It was found that hydrated hemp seed protein in a ratio of 1:1 had the best parameters for introduction into minced meat systems. It was determined that the introduction of proteins from hemp seeds (hydromodule 1:1) in the amount of 12–16 % increases the water-binding capacity of minced meat to 97.8 %, plasticity by 16.19–23.85 %, and the content of total moisture in samples of cooked sausages after heat treatment by 5.08–7.08 %, yield of the finished product up to 129.83 %. Replacing second-grade beef in cooked sausage with protein from hemp seeds causes an increase in the mass fraction of protein in finished products by 14.99–19.98 %, and mineral substances by 68.52–97.22 %. The organoleptic indicators of the experimental cooked sausages met the regulatory requirements. The safety of the developed products was established according to microbiological indicators. It has been proven that the use of hydrated protein from hemp seeds in the technology of cooked sausages in the amount of 12–14 % makes it possible to obtain a meat product with a high level of functional and technological indicators of model meat systems and organoleptic indicators of finished products. The use of protein from hemp seeds in the production of cooked sausages makes it possible to expand the range of products popular among the population

Antitumor activity of protein fraction of Jatropha curcas In DMBA and UV B induced skin tumors in mice: Expression of Bcl-2

Antitumor activity of protein fraction of Jatropha curcas In DMBA and UV B induced skin tumors in mice: Expression of Bcl-2

Mechanistic insights into the foaming properties of potato proteins based on protein composition, colloidal state, and air-water interfacial properties

Potato proteins have been shown to be suitable alternatives for animal proteins in food foam formulations but their ability to form and stabilize foam is mechanistically ill-understood. The present study aimed to better understand the foaming properties of a protein fraction isolated from potato trimmings, a by-product from the potato industry. This was achieved by comparing the properties of these in-house isolated proteins to those of commercial and highly purified potato proteins. Protein isolates (53.8 ± 4.7 g/100 g dry weight) were produced from trimmings by alkaline solubilization (pH 9.0) and isoelectric precipitation (pH 4.0). The foaming properties of the in-house isolate at pH 3.0 and 7.0 were better than those of a blend of commercial potato proteins with similar relative amounts of patatin and protease inhibitors as the in-house isolate. Notably, this was the case despite the fact that more proteins were insoluble in the in-house isolate than in the commercial potato protein suspensions. Patatin and protease inhibitors were further shown to both contribute considerably to the formation and stabilization of foams. Interestingly, a beneficial effect on foam stability was found when patatin and protease inhibitors of various colloidal sizes were present. Additionally, impurities in the in-house isolates likely contributed to their superior foaming properties. In conclusion, a protein-rich fraction with good foaming properties can be recovered from potato trimmings. Mechanistic insights were gained into the foaming properties of potato protein (mixtures). This research also highlighted the importance of assessing protein colloidal state when investigating plant protein structure-function relationships.

Ultrasound as Green Technology for the Valorization of Pumpkin Leaves: Intensification of Protein Recovery.

The recovery of valuable nutritional compounds, like proteins, from waste streams and by-products is a key strategy for enhancing production sustainability and opening up new market potential. This research aimed to use high-intensity ultrasound as an innovative technique to extract the soluble proteins from the pumpkin leaves. The impact of various sonication amplitudes and duration periods on protein yield, functional properties, antioxidant qualities, and structural characteristics, were studied. Utilization of ultrasound technology significantly increased the yield of pumpkin leaf protein by up to 40%-six times higher than maceration. The ultrasound extraction provided a RuBisCO-rich protein fraction with high radical scavenging and chelating activities, especially at 40% amplitude. Cavitation modified the tertiary and secondary structures of leaf proteins: the amount of α-helix changed based on amplitude (12.3-37.7%), the amount of random coil increased to 20.4%, and the amount of β-turn reduced from 31 to 18.6%. The alteration of the protein fluorescence spectrum (blue shift in spectrum) provides further evidence that ultrasound alters the proteins' molecular structure in comparation with maceration; the maximum tryptophan fluorescence intensity decreased from 22.000 to 17.096. The hydrophobicity values of 76.8-101.5 were substantially higher than the maceration value of 53.4, indicating that ultrasound improved the hydrophobicity of protein surfaces. Ultrasound resulted in a significant increase in solubility in an acidic environment with the increase in sonication amplitude. A 2.4-fold increase in solubility at pH 2 becomes apparent (20% amplitude; 43.1%) versus maceration (18.2%). The emulsifying ability decreases from 6.62 to 5.13 m2/g once the sonication amplitude increases by 20-70%. By combining the ultrasound periods and amplitudes, it is possible to create high-value protein leaf extracts with improved properties which can find real application as food additives and dietary supplements.

Microeukaryote metabolism across the western North Atlantic Ocean revealed through autonomous underwater profiling

Microeukaryotes are key contributors to marine carbon cycling. Their physiology, ecology, and interactions with the chemical environment are poorly understood in offshore ecosystems, and especially in the deep ocean. Using the Autonomous Underwater Vehicle Clio, microbial communities along a 1050 km transect in the western North Atlantic Ocean were surveyed at 10–200 m vertical depth increments to capture metabolic signatures spanning oligotrophic, continental margin, and productive coastal ecosystems. Microeukaryotes were examined using a paired metatranscriptomic and metaproteomic approach. Here we show a diverse surface assemblage consisting of stramenopiles, dinoflagellates and ciliates represented in both the transcript and protein fractions, with foraminifera, radiolaria, picozoa, and discoba proteins enriched at >200 m, and fungal proteins emerging in waters >3000 m. In the broad microeukaryote community, nitrogen stress biomarkers were found at coastal sites, with phosphorus stress biomarkers offshore. This multi-omics dataset broadens our understanding of how microeukaryotic taxa and their functional processes are structured along environmental gradients of temperature, light, and nutrients.

Simultaneous extraction of oil, protein and polysaccharide from Idesia polycarpa Maxim cake meal using ultrasound combined with three phase partitioning

This study explored the potential of ultrasonic-assisted three-phase partitioning (UTPP) to simultaneously extract lipids, proteins, and polysaccharides from Idesia polycarpa Maxim (IPM) cake meal, a significant byproduct of oil extraction. The impact of variables such as inorganic salt type, solid–liquid ratio, salt concentration, pH, ultrasonic time, temperature, and volume of dimethyl carbonate was examined. Based on the single-factor tests and response surface methodology (RSM), optimal conditions were identified as 30 % ammonium citrate, a 1:26 solid–liquid ratio, pH 3, 31 min of ultrasonic time, 30 °C temperature, and 15 mL of dimethyl carbonate. These conditions achieved extraction rates of 8.10 % for lipids, 5.03 % for proteins, and 10.03 % for polysaccharides, with recovery rates of 91.62 %, 83.08 %, and 93.95 % respectively. Chemical analysis showed the lipid fraction rich in linoleic acid, and the protein fraction high in glutamic acid, aspartate, and serine. The polysaccharide fraction, mainly RG-I pectin with a molecular weight of 226.58 kDa, exhibited strong thermal stability and inhibitory effects on α-glucosidase and glycation, suggesting potential for functional food and dietary supplement applications. This highlights UTPP as a sustainable method for effectively utilizing valuable compounds from IPM cake meal, outperforming traditional extraction techniques.

Can immunocrit be used as a monitoring tool for swine vaccination and infection studies?

BackgroundThe immunocrit is a cost-effective and straightforward technique traditionally used to assess passive immunity transfer to newborn piglets. However, it has not been previously used for monitoring the effect of vaccination and/or infections. Therefore, this study aimed to evaluate the usefulness of the immunocrit technique as an immunological monitoring tool in a vaccination and challenge scenario, using porcine circovirus 2 (PCV-2) as pathogen model. The immunocrit ratio was monitored in PCV-2 vaccinated (V) and non-vaccinated (NV) 3-week-old piglets (study day 0, SD0) that were subsequently challenged with this virus at SD21 and followed up to SD42. Additional techniques (PCV-2 IgG ELISA, optical refractometry, and proteinogram) were performed to further characterize the results of the immunocrit analysis.ResultsImmunocrit, γ-globulin concentration and PCV-2 S/P values followed similar dynamics: descending after PCV-2 vaccination but ascending after an experimental PCV-2 inoculation. However, statistically significant differences between V and NV animals were only found with the PCV-2 ELISA. In this case, V animals had significantly higher (p < 0.05) S/P values (S/P ratio = 0.74) than NV (S/P ratio = 0.39) pigs only after challenge at SD42. On the other hand, serum total protein obtained by refractometer (STPr) were maintained from SD0 to SD21 and increased in both groups from SD21 to SD42. Correlations between techniques were low to moderate, being the most robust ones found between immunocrit and optical refractometry (ρ = 0.41) and immunocrit with γ-globulins (ρ = 0.39). In a subset of sera, the proteinogram technique was applied to the whole serum and the supernatant of the immunocrit, with the objective to characterize indirectly the immunocrit fraction. The latter one included all protein types detectable through the proteinogram, with percentages varying between 64.3% (γ-globulins) and 82% (β-globulins).ConclusionThe immunocrit technique represented a fraction of the total serum proteins, with low to moderate correlation with all the complementary techniques measured in this study. Its determination at different time points did not allow monitoring the effect of vaccination and/or infection using PCV-2 as a pathogen model.

Corn silk as an underutilized sustainable source of plant proteins: Extraction, fractionation, structure, and techno-functional properties

Plant proteins extracted from agro-industrial waste are gaining importance in the food industry due to environmental, sustainability, and economic reasons. These proteins can be extracted and then converted into high-value ingredients, thereby promoting a circular economy. Corn silk, often regarded as an underutilized waste stream, currently has limited applications beyond its uses as a traditional medicine in some regions of the world. The present study characterized the structural, physicochemical, thermal, techno-functional, and in vitro digestibility properties of corn silk protein (CSP) and its three major fractions (albumin, globulin, and glutelin). Glutelin (67.5%) was the major protein fraction of CSP, followed by albumin (23.2%) and globulin (9.2%). A purity greater than 60% could be achieved for all fractions. The structural and techno-functional properties of the different protein fractions were impacted by their composition. The glutelin fractions had higher foaming and thermal properties but lower emulsification properties than the other fractions. However, the albumin fraction had higher in vitro digestibility and better nutritional quality. Nevertheless, the CSPs did have lysine levels below the recommended values. These findings extend the current understanding of the structural and functional properties of corn silk protein and its fractions, thereby expanding their potential applications in the food and other industries.

Lamin A/C facilitates DNA damage response by modulating ATM signaling and homologous recombination pathways

ABSTRACT Lamin A/C, a core component of the nuclear lamina, forms a mesh-like structure beneath the inner nuclear membrane. While its structural role is well-studied, its involvement in DNA metabolism remains unclear. We conducted sequential protein fractionation to determine the subcellular localization of early DNA damage response (DDR) proteins. Our findings indicate that most DDR proteins, including ATM and the MRE11-RAD50-NBS1 (MRN) complex, are present in the nuclease – and high salt-resistant pellet fraction. Notably, ATM and MRN remain stably associated with these structures throughout the cell cycle, independent of ionizing radiation (IR)-induced DNA damage. Although Lamin A/C interacts with ATM and MRN, its depletion does not disrupt their association with nuclease-resistant structures. However, it impairs the IR-enhanced association of ATM with the nuclear matrix and ATM-mediated DDR signaling, as well as the interaction between ATM and MRN. This disruption impedes the recruitment of MRE11 to damaged DNA and the association of damaged DNA with the nuclear matrix. Additionally, Lamin A/C depletion results in reduced protein levels of CtIP and RAD51, which is mediated by transcriptional regulation. This, in turn, impairs the efficiency of homologous recombination (HR). Our findings indicate that Lamin A/C plays a pivotal role in DNA damage repair (DDR) by orchestrating ATM-mediated signaling, maintaining HR protein levels, and ensuring efficient DNA repair processes.

Multiscale study of structure formation in high moisture extruded plant protein biopolymer mixes

While high moisture extrusion (HME) of plant proteins is known to result in anisotropic structures, there are still gaps in understanding the principles underlying structure formation, as well as how to study the interactions between biopolymers in mixed systems. This study aims to employ advanced rheological and microstructural tools to better characterize the structure of various HME systems and to derive some general principles for their structure formation. Pea protein isolate, with and without starch, and less refined protein fractions from pea and faba bean were used to produce extrudates with varying structures. Mechanical properties were assessed using Texture Profile Analysis, Dynamic Mechanical Analysis (DMA), and oscillatory rheology, while microstructural insights were obtained through Confocal Laser Scanning and Confocal Raman Microscopy. The addition of maize starch resulted in softer extrudates with clear partitions between protein and starch domains. The different formulations used in this study also suggest the important role of different complex carbohydrates in structure formation. Distinct parabolic shapes were observed in the extrudates, suggesting that lamellae formed during cooling, influenced by melt viscosity and the laminar flow profile resulting from the temperature gradient across the cooling die. DMA was particularly useful in evaluating differences in mechanical behavior and anisotropy. Raman microscopy proved to be valuable in assessing spatial distribution of various components and providing insights of water mobility related to competition. Evaluating various biopolymer mixes allowed for a more general view into the mechanisms of formation of structure in HME, important to enhance the development of plant-based foods.

Protein Characterization and Amino Acid Profiles of Various Wheat Varieties and Their Effect on Pasting Properties of Starch

AbstractThe present study is to investigate amino acid compositions and protein characteristics of gluten extracted from different varieties of wheat and their influence on the pasting properties of starch. The starch and gluten are isolated separately from 12 wheat varieties categorized by grain hardness, i.e., hard (HW), medium hard (MHW), and soft (SW)with value ranged from 83 to 95, 72 to 80, and 17 to 29, respectively. Amino acid profiling reveals significant variations among the wheat varieties, with extraordinarily soft wheat (Ex‐SW) varieties exhibited higher levels of essential amino acid including methionine, phenylalanine, tryptophan, threonine, isoleucine, histidine, lysine, and valine. Protein fraction analysis indicates that gluten from HW varieties has a higher proportion of gliadin and lower glutenin content compared to MHW and Ex‐SW varieties. Furthermore, starch from Ex‐SW varieties possesses higher pasting parameters than MHW and HW varieties. Further, incorporation of gluten significantly reduced the pasting characteristics of starch. Flours with higher protein content demonstrate lower paste breakdown, highlighting the influence of protein content on viscoelastic properties. Furthermore, principal component analysis (PCA) reveals associations between specific amino acids and protein fractions with various pasting properties, highlighting the complex interplay between gluten composition and starch behavior.

Antithrombotic and anticariogenic activity of peptide fractions from cowpea (Vigna unguiculata) protein hydrolysates.

Protein-derived peptide fractions can play a key role in the physiological and metabolic regulation and modulation of the body, which suggests that they could be used as functional ingredients to improve health and to reduce the risk of disease. This work aimed to evaluate the in vitro antithrombotic and anticariogenic bioactivity of hydrolysates and protein fractions obtained from cowpea (Vigna unguiculata) by biocatalysis. Cowpea protein concentrate was hydrolyzed by sequential action with two enzyme systems, Pepsin-Pancreatin or Alcalase-Flavourzyme. There was extensive enzymatic hydrolysis, with degrees of hydrolysis of 34.94% and 81.43% for Pepsin-Pancreatin and Alcalase-Flavourzyme, respectively. The degree of hydrolysis for the control treatments, without the addition of the enzymes Pepsin-Pancreatin and Alcalase-Flavourzyme was 1.1% and 1.2%, respectively. The hydrolysates were subjected to fractionation by ultrafiltration, with five cut-off points according to molecular weight (<1, 1-3, 3-5, 5-10 and >10 kDa). The Alcalase-Flavourzyme hydrolysate led to 100% inhibition of platelet aggregation, while the Pepsin-Pancreatin hydrolysate showed 77.41% inhibition, but this was approximately 100% in the ultrafiltered fractions. The highest anticariogenic activity was obtained with the Pepsin-Pancreatin system, with 61.55% and 56.07% for calcium and phosphorus demineralization, respectively. Hydrolysates and their peptide fractions from Vigna unguiculata exhibited inhibition of platelet aggregation and protection of tooth enamel and have the potential for use in the development of functional products with beneficial health effects. © 2024 Society of Chemical Industry.

Production of Bioactive Peptides from Microalgae and Their Biological Properties Related to Cardiovascular Disease

Microalgae are a substantial group of unicellular prokaryotic and eukaryotic marine organisms. Due to their high protein content of 50–70%, microalgae have the potential to become a sustainable alternative protein source, as well as aiding in the development of bioactive peptide-based nutraceuticals. A series of major steps are involved in the production of peptides from microalgae, which include the disruption of the microalgal cell wall, the hydrolysis of proteins, and the extraction or isolation of peptides derived from hydrolysis. Physical methods of cell wall disruptions are favored due to the ability to obtain high-quality protein fractions for peptide production. Bioactive peptides are protein fragments of two to twenty amino acid residues that have a beneficial impact on the physiological functions or conditions of human health. Strong scientific evidence exists for the in vitro antioxidant, antihypertensive, and anti-atherosclerotic properties of microalgal peptides. This review is aimed at summarizing the methods of producing microalgal peptides, and their role and mechanisms in improving cardiovascular health. The review reveals that the validation of the physiological benefits of the microalgal peptides in relation to cardiovascular disease, using human clinical trials, is required.

Exotoxins secreted by Clostridium septicum induce macrophage death: Implications for bacterial immune evasion mechanisms at infection sites

The induction of macrophage death is considered a potential mechanism by which components secreted by Clostridium septicum are used to evade the innate immune response and cause tissue damage. This study aimed to determine the effects of partially purified fractions of extracellular proteins secreted by C. septicum on the death of mouse peritoneal macrophages. Elicited mouse peritoneal macrophages were incubated with partially purified fractions of proteins secreted by C. septicum into the culture medium. After incubation, the protein fraction with a molecular weight ≥100 kDa caused significant cell death in macrophages, altered cell morphology, increased the expression of markers of apoptosis and autophagy, and increased the expression (protein and mRNA) of IL-10 and TNFα. Our data suggest that the proteins secreted by C. septicum (MW, ≥100 kDa) induce cell death in macrophages by promoting autophagy-triggered apoptosis. This study may contribute to our understanding of the molecular mechanism of immune evasion by C. septicum at the infection site.

Advancements in Pulse Starches: Exploring Non-Thermal Modification Methods.

The surge in the global demand for plant-based proteins has catapulted pulse protein into the spotlight. To ensure economic viability and sustainable production, it is crucial to utilize pulse starch, a by-product of plant protein fractionation. Despite the increasing interest in pulse starches, there is a notable gap in knowledge regarding their modifications and applications compared to cereal and tuber starches. Non-thermal techniques such as electron beam radiation, static high pressure, microfluidization, and cold plasma are emerging as innovative methods for starch modification. These techniques offer significant advantages, including enhanced safety, environmental sustainability, and the development of unique functional properties unattainable through conventional methods. However, challenges such as equipment availability, high costs, and energy consumption hinder their widespread adoption. In light of the growing emphasis on "clean and green labelling" and effective "waste management" in food production, evaluating non-thermal techniques for pulse starch modification is critical. This review aims to thoroughly assess these non-thermal techniques and their combinations, offering valuable insights for researchers and the food industry. By maximizing the potential of pulse starches in innovative food applications, it provides a comprehensive guide for effective non-thermal methods that add value and align with sustainable practices.

The regulatory effects of water probiotic supplementation on the blood physiology, reproductive performance, and its related genes in Red Tilapia (Oreochromis niloticus X O. mossambicus)

Probiotics are becoming increasingly popular as eco-friendly alternatives in aquaculture. However, there is limited research on their impacts on the reproductive efficiency of Red Tilapia (Oreochromis niloticus x O. mossambicus) broodstock. Therefore, this experiment aimed to explore the combined effects of selective probiotics Bacillus subtilis and B. licheniformis (BSL; 1:1) added to water on blood hematology, serum metabolites, gonadal histology, reproductive performance, and reproductive associated genes in Red Tilapia broodstock. Tilapia broodfish weighing 140–160 g were stocked in four treatment groups: control (T0), and the other three groups were added different levels of BSL to the water as follows: T1 (0.01 g/m3), T2 (0.02 g/m3), and T3 (0.03 g/m3), respectively. Results indicate that BSL administration significantly improved RBCs, hemoglobin, hematocrit, MCH, and MCHC, with the highest improvement seen in the T3 group (P < 0.05). BSL added to the fish water significantly enhanced serum protein fractions (total protein, albumin, and globulins), while AST, ALT, ALP, creatinine, uric acid, and glucose were significantly diminished in a dose-dependent way (P < 0.05). Adding 0.02–0.03 g/ m3 of BSL resulted in higher antioxidant status (superoxide dismutase and catalase) compared to other groups (P < 0.05). Testosterone levels were higher in T3 than in other groups (P < 0.05). All female hormones (LH, FSH, estradiol, and progesterone) were substantially augmented by the addition of BSL. Additionally, the BSL groups exhibited higher GSI, HSI, VSI (male only), egg diameter (mm), mean number of fry/fish, and mean fry weight (g) compared to the control group (P < 0.05). Expression of reproductive-associated genes (vasa, nanos1a, nanos2, dnd1, pum1, AMH, and vtg) were significantly up-regulated in the gonads of fish in the 0.03 g/m3 treatment. The histological gonadal structure exhibited that BSL improved gonad maturation in both genders of Tilapia fish. Overall, adding a mixture of B. subtilis and B. licheniformis (0.03 g/m3 water) can accelerate reproductive performance in Red Tilapia through up-regulation of reproductive genes and enhance the health profile.