Chubitchi, a traditional brewed alcoholic beverage from Meghalaya, was prepared under controlled laboratory conditions. A comprehensive storage study was conducted over four weeks under refrigerated conditions to examine its physicochemical properties (pH, acidity, alcohol content) and bio-functional attributes (antimicrobial, antioxidative, ACE inhibitory, anti-diabetic activities, and phenolic content). Over the four weeks, the pH decreased to 3.31, while acidity increased to 2.03%. Ethanol content peaked at 5.62% during week 2. Lactic acid bacteria and yeast counts declined to 5.21 log CFU/mL and 4.86 +/- 0.19 log CFU/mL, respectively, by week 4. Chubitchi demonstrated antimicrobial activity against K. pneumoniae, E. coli, and S. aureus. Antioxidative activity peaked in weeks 3 (86.19 +/- 0.00%) and 4 (86.04 +/- 0.06%). ACE inhibition increased non-significantly from 17.03 +/- 0.28% to 23.10 +/- 0.92%. Anti-diabetic potential peaked at 3.53% (alpha-amylase inhibition), 20.42% (alpha-glucosidase inhibition), and 39.50% (lipase inhibition). Total phenolic content rose from 0.22 mg/mL to 0.27 mg/mL. Organoleptic evaluation revealed declining sensory scores through day 40. Furthermore, a 1:8 dilution of Chubitchi was found to be non-cytotoxic in a murine macrophage cell line, exhibiting significant antiinflammatory activity and reduced reactive oxygen species (ROS) production. This study provides critical insights into physicochemical and biofunctional dynamics of Chubitchi during storage, supporting its potential as a functional beverage.

This study addresses the growing demand for natural functional foods with therapeutic potential, particularly for managing diabetes, oxidative stress, and inflammation. The aim of this research was to investigate the biofunctional attributes of camel milk fermented with Limosilactobacillus fermentum (KGL4) and Saccharomyces cerevisiae (WBS2A), with a focus on its antidiabetic, antioxidative, proteolytic, anti-inflammatory, and cytoprotective activities. Fermentation was performed for 0, 12, 36 and 48 h at 30 °C (2% inoculum rate). Fermented camel milk significantly enhanced α-amylase (81.33%) and α-glucosidase (68.37%) inhibition, demonstrating strong antidiabetic potential. Antioxidant activity, as assessed through the ABTS assay, progressively increased with incubation time, reaching a peak of 65.87% at 48 h. Proteolytic activity also rose significantly, attaining a maximum of 6.41 mg/ml (2.5% inoculum concentration at 30 °C/48 h). Chromatographic profiling via RP-HPLC revealed increased bioactivity in 3 kDa permeate (antidiabetic) and 10 kDa retentate (antioxidant) samples, suggesting the role of low-molecular-weight peptides. 2D gel electrophoresis and SDS–PAGE confirmed proteolytic cleavage, revealing the presence of smaller peptide fragments in the fermented samples than in the unfermented controls. Further structural analysis via FTIR and confocal laser scanning microscopy (CLSM) demonstrated secondary structure modifications, including increased β-sheet formation and reduced aggregate size. Molecular docking studies revealed that the identified peptide sequence CCFSSCAMR effectively bound to the human digestive enzymes hBAL, hPAM, and hMGA via hydrogen bonding and hydrophobic interactions, supporting its potential inhibitory function. Additionally, fermented CM displayed strong anti-inflammatory and cytoprotective effects on LPS-stimulated RAW 264.7 macrophages. This study highlights the potential of fermented camel milk as a value-added functional food with significant antidiabetic, antioxidant, anti-inflammatory, and cytoprotective properties. The generation of low-molecular-weight bioactive peptides through targeted microbial fermentation provides a scientific foundation for developing natural, nonpharmacological interventions for metabolic and inflammation-related disorders. These findings support the application of fermented camel milk in functional food and nutraceutical development.

Carbapenem-resistant Pseudomonas aeruginosa (CRPA) is recognized as a global health problem due to its antibiotic resistance and zoonotic potential, posing serious risks in both clinical and dairy settings. This study analysed the phenotypic and genotypic characteristics of Pseudomonas spp. isolated from bovine mastitis milk collected from cattle farm in Palanpur, Gujarat. A total of 60 milk samples were tested, and 43 isolates were identified as Pseudomonas spp. using Brain Heart Infusion agar and MALDI-TOF MS, followed by antibiotic susceptibility testing. Ten of these isolates demonstrated extensive drug resistance (XDR), whereas two exhibited pan-drug resistances. Ampicillin (100 %), amoxicillin-clavulanate (90 %), cefoxitin (90 %), and tetracycline (80 %) all showed considerable resistance, whilst aztreonam and amikacin showed the highest sensitivity. Wholegenome sequencing (WGS) of both prevalent carbapenem-resistant isolates, Pseudomonas aeruginosa SKN681 and Pseudomonas otitidis SKN246, revealed genome sizes of 8.08 Mb and 5.91 Mb, with 9043 and 5595 coding sequences, respectively. Genomic research revealed several beta-lactamase genes, multidrug efflux systems (MexAB-OprM, EmrAB-OMF, and OprD), virulence determinants, integrases, and transposases, demonstrating considerable genomic flexibility and horizontal gene transfer potential. Phylogenetic research verified their close relationship to high-risk Pseudomonas lineages. Overall, this study provides the preliminary comprehensive genomic analysis of carbapenem-resistant Pseudomonas spp. from bovine mastitis in Gujarat, revealing their multidrug-and pan-drug-resistant nature, diverse AMR gene repertoire, and zoonotic potential, emphasizing the critical need for improved antimicrobial resistance surveillance, responsible antibiotic stewardship, a One Health-based approach to mitigate their transmission of these diseases in both animal and human populations.

This study explored the antioxidative and antidiabetic potential of bioactive peptides derived from Panchali sheep milk fermented with Lacticaseibacillus rhamnosus (M9, MTCC 25516) and Saccharomyces cerevisiae (WBS2A, MG101828). Fermentation was conducted at 30°C, and measurements were taken at 0, 12, 24, 36, and 48 h. Maximum activity was observed at 48 h, including antioxidant activity (ABTS free radical scavenging) of 28.62%, and α-glucosidase and α-amylase inhibition of 71.19% and 65.90%, respectively. Proteolysis was optimized using varying inoculum levels and incubation times, with maximal activity (6.31 mg/mL) attained at 2.5% inoculum after 48 h. Fermented sheep milk notably suppressed LPS-induced cytotoxin and ROS production in RAW 267.4 cells, suggesting anti-inflammatory effects. Structural and molecular changes in proteins were validated using CLSM and FTIR, revealing alterations in the protein networks. SDS-PAGE revealed prominent protein bands between 10-85 kDa, while no bands were detected in permeate samples. In contrast, 2D gel electrophoresis localized 35 distinct protein spots within the 15–70 kDa molecular weight range. Water-soluble extracts were further analyzed using RP-HPLC and MASCOT software, with sequences validated through the BIOPEP database for antioxidant and antidiabetic activity. Notably, molecular docking of the ITMPLW peptide showed a high binding affinity for digestive enzymes, confirming its multifunctional bioactivity. Overall, fermented Panchali sheep milk is a novel source of peptides with strong antioxidant and antidiabetic potential.

This investigation was conducted to analyse the antioxidant activity, proteolytic activity and ability of short-chain fatty acids (SCFAs) production from fermented cheddar cheese whey fractions inoculated with Limosilactobacillus fermentum, MF951096 (M4) @ 2.5% when incubated at 37 degrees C for 48 h. Ultrafiltration membranes with appropriate molecular weight thresholds were used to isolate low-molecular-weight peptides from fermented whey, including the 3/ kDa permeate, 10/ kDa permeate, and retentate fractions. In acidity and pH profile, there was no significant difference observed. Significantly higher peptide content (7.84mg/mL) and antioxidant activity (65.81%) were observed in the 10/ kDa retentate fraction; However, the 10 kDa retentate showed a lactic count among all the fractions. Also we characterized the peptide fractions by analysing their chromatographic peaks using Reverse-Phase High-Performance Liquid Chromatography. The 10/ kDa retentate fraction produced the most SCFA i.e. 46.68 & igrave;g/mL acetic acid,21.59 & igrave;g/mL propionic acid, and 5.27 & igrave;g/mL butyric acid, respectively. From this investigation, we concluded that the conversion of dairy waste into bioactive molecules presents a novel approach with conceivable applications in the dairy, food and pharmaceutical sector.

Acinetobacter spp. are opportunistic bacteria that are increasingly found in raw milk in dairy farms, posing a public health risk due to their propensity to acquire and transmit antimicrobial resistance (AMR). The purpose of this study was to characterize Acinetobacter isolates from dairy cattle raw milk and milk products in Anand, Gujarat, India, using both phenotypic and whole-genome sequencing (WGS) approaches. A total of 120 raw milk and milk product samples were collected and analysed, from which 20 samples exhibiting Acinetobacter spp were 80 % MDR (Multidrug resistance) While two Extensively drug Resistance (XDR) These isolates were identified using standard microbiological and biochemical techniques, revealing oxidase-negative, catalase-positive, and non-fermentative profile. Species-level identification using MALDI-TOF mass spectrometry confirmed the isolates as Acinetobacter schindleri SKN291 and Acinetobacter indicus SKN436. Both isolates were further analysed through whole-genome sequencing, and phenotypic analysis revealed that a high proportion of the Acinetobacter spp. isolates were extensively drug-resistant (XDR). According to CLSI recommendations, phenotypic antimicrobial susceptibility testing demonstrated resistance to numerous types of antibiotics but still showed susceptibility to important last-resort medications like colistin and imipenem. WGS of A. schindleri SKN291 and A. indicus SKN436 produced high-quality genome assemblies (3.20 Mb and 2.85 Mb, respectively). These assemblies showed a wide range of AMR genes, including ones that make bacteria resistant to beta-lactams, aminoglycosides, quinolones, and tetracyclines, as well as mobile genetic elements (MGEs) that facilitate horizontal gene transfer. Both isolates carried efflux pumps (MacA, MacB, MdfA/Cmr, TolC/OpmH), multiple antibiotic target site genes, and target replacement proteins (FabG, HtdX), with SKN291 lacking gidB and SKN436 harboring additional determinants including OXA-134, KatG, BcrC, GdpD, PgsA, and the regulator OxyR. Phylogenetic research validated the correct taxonomic classification and indicated evolutionary links with other Acinetobacter species. The identification of multidrugand extensively drug-resistant bacteria in the dairy environment emphasizes the necessity of genomic surveillance, enhanced milk handling hygiene, and sensible antimicrobial usage in mitigating AMR concerns within a One Health framework.

Kefir is a probiotic-rich beverage that is fermented with beneficial microorganisms that live together in symbiotic association within kefir. The most established health benefits of kefir include gut and digestive health, owing to its rich probiotic diversity, which enhances intestinal balance and lactose digestion. Kefir consumption is associated with certain health enhancing outcomes, including anti-atherosclerotic, anti-inflammatory, antihypertensive, antioxidant, antibacterial, anticancer, and antidiabetic effects. Strong outputs were obtained via antimicrobial, immunomodulatory, anti-inflammatory, and antioxidant effects, supported by extensive experimental evidence. Emerging studies suggest anticancer, antidiabetic, and neuroprotectiveepotentials, although further clinical validation is required. Using the Scopus database, 202 documents published between 2013 and 2022 were identified and submitted for bibliometric analysis using VOSviewer software. Regarding recent applications, 107 documents on kefir published between 2021 and June 2023 were identified. This systematic review provides updated insights into the evaluation of kefir, including its nutritional and microbiological compositions. It also highlights the biological activities of kefir microbiota and emphasizes the significance of kefiran as a functional component.

Fermented camel milk is gaining attention as a functional food due to the formation of bioactive peptides during microbial fermentation. This study aimed to investigate the generation of bioactive peptides and their associated antidiabetic, antioxidant, and anti-inflammatory activities in camel milk fermented using Lacticaseibacillus rhamnosus M9 and Saccharomyces cerevisiae WBS2A. The in vitro fermentation process was evaluated in relation to proteolytic activity and peptide formation under varying fermentation times and inoculum concentrations. Antidiabetic activity was assessed using α-amylase and α-glucosidase inhibition assays, while antioxidant potential was determined using the ABTS assay. Peptide profiles and metabolite changes were characterized using RP-HPLC, SDS-PAGE, and 2D electrophoresis, and structural modifications were analysed through FTIR and CLSM. Fermentation significantly enhanced proteolytic activity, leading to the release of low-molecular-weight peptides (3–10 kDa) associated with improved bioactivity. Fermented samples exhibited notable α-amylase (70.68

Cactus pear popularly known as prickly pear, in the local languages of India, possess many medicinal values. Traditionally this fruit has been utilized as therapeutic agent across various parts of the world to treat conditions such as asthma, anemia, wound infections, intestinal inflammation, and diabetes. It is valuable fruit because it contains important phytochemicals that promote human health. The nutritional profiling of the fruit extract revealed the presence of polyphenols, pigments, flavonoids, vitamins, carbohydrates, minerals, fibers and water content, all of which offer potential health benefits. Therefore, considering the cactus pear fruit's nutritional quality, medicinal values and potential for its application in diverse food products development, this review highlights the composition of cactus pear fruit, medicinal properties and utilizations in the production of a wide range of food items, including juices, beverages, jams, jellies, sirups, concentrates, natural colorants, functional dairy-based products, and fermented foods.

Interest in probiotics and its products has grown recently among consumers and researchers alike. The current concept aims to create a non-fermented milk product preparation that can be consumed at every meal. It can be taken after lunch or dinner as a dessert or savoury supplement. Such product will fully satisfy the body's minimal needs by supplying ascorbic acid (AA) and a daily dose of probiotic bacteria in encapsulated form. The concept of such product development was standardized and optimized to have sufficient isabgol, a prebiotic component to provide the host with more dietary content. The present investigation a recipe for preparation of synbiotic milk pudding. The current study focuses on optimizing the technique for making synbiotic milk pudding. In this study, addition rate of sugar, agar-agar, isabgol, co-encapsulated beads were optimized and rates of addition of sugar, agar-agar, isabgol @ 16%, 1.5%, 0.5% respectively gained better sensory scores. For microcapsules, it was @1% that showed optimum cell viability and ascorbic acid concentration along with higher sensory scores. The finalized product had a probiotic viability of 9.457 +/- 0.08 log cfu/g that is within the range of effective probiotic dosage. The product also has 38.21 +/- 0.80 mg/100 g of ascorbic acid concentration that means as per the serving size of 25 g it confers 9.55 mg of ascorbic acid that can fulfil about 11.6 % of RDA (for an average adult human being) of vitamin C. The developed synbiotic non-fermented milk pudding could be better option to have three-in-one effect of a probiotic culture, ascorbic acid and benefits of isabgol for better heath.

Ready-to-eat kiwifruit refers to kiwifruit with a uniform texture, maturity and favorable taste that can be consumed immediately after purchase without the requirement for a natural ripening process. Although extensive research has been conducted on the application of forchlorfenuron (CPPU) in kiwifruit cultivation, studies on its impact on the "edible window" of ready-to-eat kiwifruit and relevant mechanisms remain limited. In this study, to investigate the impact of CPPU treatment on the edible shelf-life qualities of ready-to-eat kiwifruit, we conducted an in-depth analysis of multiple aspects, including fruit qualities, enzyme activities, cellular structure, and metabolites. The results indicated that CPPU treatment increased the fruits' volume and weight, but decreased the firmness. During storage, CPPU treatment increased the respiratory rate of kiwifruit, with a peak respiratory intensity of 46.14 mgkg-1h-1 and 34.36 mgkg-1h-1 for the samples treated with or without CPPU, respectively. This suggested that CPPU accelerated the conversion of starch to sugar and the ripening of kiwifruit. At room temperature, the edible window of ready-to-eat kiwifruit not exposed to CPPU was 7 days, while the samples treated with CPPU was 5 days. This was probably attributed to the reduction in antioxidant enzyme activities like catalase and superoxide dismutase, and the increase in a-amylase activity and malondialdehyde content after treatment with CPPU. The present study supplies useful information for developing ready-to-eat kiwifruit, especially for CPPU-treated kiwifruit.

This study explores the antidiabetic and anti-inflammatory activities of polypeptides from fermented whey protein isolate (WPI) and soy protein isolate (SPI) using Limosilactobacillus fermentum (KGL4) MTCC 25515. Both fermented WPI and SPI demonstrated significant α-amylase and α-glucosidase inhibition, with SPI showing higher inhibition rates. Proteolytic activity peaked at 48 h and a 2.5% inoculation rate, with sodium dodecyl sulfate-polyacrylamide gel electrophoresis (PAGE) and two-dimensional (2D)-PAGE revealing changes in protein profiles and peptide distribution. Ultra-filtered fractions, particularly the 3 kDa permeates, exhibited strong antidiabetic effects. Furthermore, fermented WPI and SPI reduced nitric oxide production and pro-inflammatory cytokines in RAW 264.7 cells without causing cytotoxicity at 2 mg/mL concentrations. Confocal microscopy showed that fermentation led to the formation of smaller peptide structures, suggesting that KGL4-fermented WPI and SPI are promising sources of bioactive peptides with potential applications in functional foods and therapeutics.

The present investigation was conducted on encapsulated beads formed from extrusion and emulsion methods having a co-encapsulated probiotic culture and L-ascorbic acid (AA). These were tested against pathogens and its responses in simulated gastric conditions. The results of study for the survival of probiotic viability as well as changes in ascorbic acid concentration in simulated gastro-intestinal conditions showed that treatment wise there was a significant difference (p<0.05) and mean concentration was significantly higher for emulsion method compared to extrusion method. For antimicrobial activity, four samples were prepared-control sample (C), encapsulated culture by emulsion method (Em), extrusion method (Ex), heat treated free cells (H). Em sample showed highest inhibition zone (16.33 +/- 2.08 mm) against Enterococcus faecalis ATCC 29212 and lowest inhibition zone (12.7 +/- 0.58 mm) was observed against E. coli MTCC 1687 and Staphylococcus aureus MTCC 7373. For Ex sample, highest inhibition zone (14.00 +/- 1.0 mm) was seen against Enterococcus faecalis ATCC 29212 and lowest inhibition zone (9.07 +/- 1.15 mm) was observed against Staphylococcus aureus MTCC 7373. The beads prepared by emulsion method survived better (p<0.05) in simulated gastro intestinal conditions than beads formed by extrusion method in terms of cell viability, AA concentration and in acidic solution. Hence, emulsion method could be the better methods for protecting sensitive ingredients and such prepared co-encapsulated beads offer better option in delivering functional items to individuals via foods.

The 2024 Nobel Prize awarded for protein structure prediction has strengthened the reliability of in silico approaches for protein and peptide research. In recent years, food-derived bioactive peptides (BAPs), which are small amino acid chains produced from food proteins, have garnered increasing interest owing to their promising health benefits. However, traditional approaches to BAPs production are often time-consuming, expensive, and unpredictable. The emergence of in silico methods has transformed BAPs research by enabling high-throughput screening and strategic utilization in food and pharmaceuticals. Molecular docking approaches have considerably expedited BAPs research by predicting the binding affinities and molecular interactions with target proteins. This review explores recent developments in computational approaches to BAPs discovery, highlighting their viability and sustainability. It provides a broad overview of advances in in silico BAPs production and molecular docking methods, delves into the appraisal of bioactivity, toxicity, and allergenicity, and discusses ligand and receptor preparation and molecular simulations. Future research should focus on improving docking algorithms, integrating multiscale modeling techniques, and incorporating high-throughput experimental screening for better validation. By addressing these challenges, the in silico techniques can play an important role in the efficient identification of novel food-derived BAPs with therapeutic potential.

This study investigates the biofunctional and antioxidant properties of chia seed protein hydrolysates produced using subcritical water (SBW) hydrolysis, a novel and environmentally friendly approach, in comparison to conventional alkali hydrolysis. For the first time, SBW hydrolysis was applied to deoiled, mucilage-free chia seed flour (DCF) to obtain protein hydrolysates. The SBW-derived hydrolysates exhibited significantly higher total phenolic (1.712 +/- 0.144 mg gallic acid equivalent/g DCF) and flavonoid content (53.42 +/- 12.21 mg quercetin equivalent/g DCF) compared to those obtained via alkali hydrolysis. The effects of processing temperature and duration on hydrolysate's biofunctional properties were evaluated, with temperature identified as a crucial factor. Antioxidant activity and enzyme inhibitory assays confirmed the potential of SBW hydrolysates for use in functional food formulations. This chemical-free approach offers a sustainable alternative for protein hydrolysis, contributing to the advancement of eco-efficient food processing technologies.

Introduction: This study aims to valorise cheese whey waste by converting it into bioactive peptides that have several health benefits, potentially leading to the development of nutraceuticals and functional foods and also used in pharmaceutical industry. Methods: The study evaluates the antidiabetic, antioxidative, and anti-inflammatory properties of fermented cheese whey with Limosilactobacillus fermentum (M4), along with the production of antioxidative and antidiabetic peptides. SDS PAGE and 2D PAGE were also performed to identify proteins by molecular weight and isoelectric point, while RP-HPLC distinguished peptide fractions. Peptide sequences from 2D gel spots were identified using RPLC/MS, and RP-HPLC analyzed 3 kDa and 10 kDa permeates. Peakview software characterized the LC/MS results, and FTIR analysis measured structural changes in bioactive peptides. Results: The antioxidative and antidiabetic properties in cheese whey fermented with M4 showed a progressive growth over extended incubation periods, higher effects were observed after fermentation for 48 hours. Inhibitory activities in α-glucosidase, α-amylase & lipase were found to be 65.39%, 66.09%, and 56.74% respectively. ABTS assay was performed to measure antioxidant activity (63.39%) and the highest proteolytic activity (7.62 mg/ml) was measured at 2.5% inoculation rate for 48 hours. In SDS-PAGE, protein bands between 10 & 30 kDa were observed, whereas peptide spots within the range of 10 to 70 kDa were also visualized on the 2D PAGE. RP-HPLC was used to distinguish different fractions of a peptide. Peptide sequences from 2D gel spots were identified using RP-HPLC & RPLC/MS. Peakview software was utilized to characterize the LC/MS results. Sequences of peptides generated from α-lactalbumin and β-lactoglobulin were searched in the BIOPEP database to validate the antidiabetic and antioxidative activities of fermented cheese whey peptides. 0.50 mg/mL of fermented cheese whey significantly LPS suppressed the production of proinflammatory cytokines as well as the mediators that govern them including IL-6, IL-1β, NO, and TNF-α in RAW 264.7 cells. FTIR was used to analysis of protein secondary structure and conformational changes. Conclusion: This study aims to the production of antidiabetic and antioxidative peptides from dairy waste, and cheese whey.

Sheep milk is a rich source of proteins with potential to generate bioactive peptides through microbial fermentation. However, limited studies have focused on the Panchali sheep breed of India and the multifunctional health properties of its fermented products. This study investigated the generation of bioactive peptides with antidiabetic, antioxidative, and anti-inflammatory properties through the fermentation of sheep milk using a potent LAB, Limosilactobacillus fermentum (KGL4, MTCC 25515), in combination with the yeast strain Saccharomyces cerevisiae (WBS2A, MG101828). Optimal bioactivity was observed after 48 h of fermentation at 30 °C, with antioxidant activity reaching 40.08

This study was aimed at producing antioxidant and antidiabetic peptides through lactic fermentation of cheddar cheese whey for the production of valuable bioactive compounds from dairy waste. Fermented whey peptide was prepared using Lactiplantibacillus plantarum KGL3A from cheddar cheese whey inoculated with 2