A vegetable-based probiotic drink provides an alternative to dairy products for delivering beneficial microorganisms. This study was conducted to develop a fermented probiotic drink using cucumber juice. The probiotic starter culture used was Lacticaseibacillus casei NCDC-17. The fermentation process was optimized to enhance antioxidant activity, total phenolic content, and microbial viability. A rotatable central composite design using response surface methodology was used to set up the experimental design. The independent variables for the optimization were incubation temperature (30-45 °C), pH (5-9), and inoculum size (2-5%). The optimal fermentation conditions were observed to be incubation temperature 37.5 °C, pH 7, and inoculum size 3.5%. The results showed that the pH, total soluble solids, total flavonoids, and total sugar content were decreased after the fermentation. It was observed that the total phenolic content, and antioxidant activity were increased to a significant level after the fermentation. Results revealed that the fermented drink inhibits the key enzymes (alpha-amylase, and alpha-glucosidase) related to diabetes. The volatile compounds were identified using Gas Chromatography-Mass Spectrometry. A total of 54 distinct volatile compounds were identified following the fermentation process, indicating significant biochemical transformation and metabolic activity of the microbial culture. Cell-free supernatant of the drink exhibited antimicrobial activity with the highest and lowest inhibition against Escherichia coli (14 mm), and Staphylococcus aureus (11 mm), respectively. The fermented sample was evaluated for its cytotoxic potential against MCF-7 human breast cancer cell lines. The fermented sample exhibited marked growth inhibitory effects, against MCF-7 cell lines. Sensory evaluation was performed using a 7-point hedonic scale to evaluate attributes such as appearance and colour, mouth feel, aroma and flavor, taste, and overall acceptability. Results showed that the probiotic drink was well accepted.

The development of plant-based yogurt alternatives continues to expand in response to lactose intolerance, sustainability concerns, and demand for nondairy fermented products. Young coconut drink, derived from a mixture of coconut water and young coconut flesh, has been reported to contain fermentable sugars, minerals, and bioactive compounds that support microbial growth, suggesting its potential as a fermentation substrate. However, its fermentation characteristics remain underexplored. This study evaluated the effects of starter culture concentration (0.8% and 1%) and fermentation time (8 and 10 h) on the microbiological, physicochemical, nutritional, mineral, and sensory properties of young coconut drink-based yogurt. Yogurt was produced using Lactobacillus delbrueckii subsp. bulgaricus and Streptococcus thermophilus at 45°C. All formulations achieved lactic acid bacteria viability above 107 CFU/g, indicating adequate LAB viability after fermentation. Starter concentration significantly influenced acidification, viscosity, proximate composition, carbohydrate utilization, and sensory acceptability, whereas fermentation time exerted a secondary effect. The formulation containing 1% starter fermented for 10 h exhibited the most balanced profile, combining desirable texture, acidity, and overall acceptability. This selected formulation showed a fatty acid profile dominated by medium-chain fatty acids, particularly lauric acid, a diverse amino acid profile characteristics of the plant-based matrix, and high mineral bioaccessibility (> 90%) for potassium, magnesium, and iron as determined by an in vitro gastrointestinal digestion model. Overall, young coconut drink-based yogurt demonstrates potential as a plant-based fermented beverage derived from locally available resources. However, further studies are required to enhance its nutritional profile and sensory characteristics.

Cow's milk and its derivatives are a food group of recognized nutritional value during childhood and adolescence, and their consumption is endorsed by the main pediatric societies from 1 year of age. However, studies indicate that a significant portion of Brazilian children and adolescents do not meet the consumption recommendations, compromising the intake of essential nutrients. This expert review critically evaluated the scientific evidence on cow's milk consumption in this age group, in a question-and-answer format, based on a search in the PubMed/MEDLINE, Scopus, Web of Science and ResearchGate databases, with a final sample of 70 publications. The results indicate that regular consumption of cow's milk is safe and beneficial for healthy children and adolescents, contributing to physical growth, bone health, favorable body composition, and potential improvement in cognitive performance. Contraindications are restricted to specific conditions, such as cow's milk protein allergy and lactose intolerance. However, some findings-particularly those related to cognitive performance-remain heterogeneous, highlighting the need for further well-designed studies. It is concluded that, in the absence of these conditions, there is no scientific basis for the exclusion of cow's milk from the diet of this population group, and its consumption should be and its consumption should be encouraged within the context of a balanced and individualized diet.

This ecological study analyzed the temporal trend of special dietary needs in children aged 2 to 12 enrolled in the public school system of Belo Horizonte (MG) between 2014 and 2023. Secondary data from medical reports filed at schools, provided by the Municipal Department of Food and Nutritional Security, were used. Variables included year, gender, age, school location and type of dietary restriction. The analysis used descriptive statistics and Prais-Winsten regression. Among the 1967 children, 12.6% had some special dietary need, especially lactose intolerance (39.8%), cow's milk protein allergy (12.6%) and egg allergy (4.2%). From 2021 onwards, there was a significant increase, with 421 cases of intolerance in 2023. The annual increase rates were: 6.69% (intolerance), 5.51% (allergies) and 5.74% (selectivity). Around 43.4% of the reports did not specify the restriction. The high level of adherence to school meals (54.6%) highlights the role of the PNAE in food safety. The findings reinforce the need for intersectoral strategies to ensure inclusive and safe food in the school environment.

Adsorption is the simplest method of enzyme immobilization, but it often requires additional treatments to improve stability. These treatments, however, may impact enzymatic activity. This study investigated the effect of crosslinking on the stability of β-galactosidase immobilized by adsorption on A-568 resin and its implications for enzymatic activity. The immobilized enzyme was crosslinked with different concentrations of glutaraldehyde (0.50% to 0.80% v/v). Parameters such as thermal stability (30°C to 60°C), stability during consecutive reuse, and storage stability for 90 days at different pH values (3-8) were evaluated. The feasibility of using the immobilized enzyme in an industrial process was also evaluated, an aspect that is not usually addressed. The conversion capacity was evaluated in a continuous reactor at different feed flow rates (4-10 mL/min). The results showed that crosslinking with glutaraldehyde (0.50%) provided greater stability for the enzyme adsorbed on the resin, which retained more than 90% of its initial activity at the end of the consecutive use cycle. Crosslinking improved the thermal stability of the enzyme, and its half-life was more than doubled at all temperatures. Storage conditions affected the non-crosslinked enzyme more, which showed intolerance to pH extremes, losing more than 60% of its initial activity at pH 3, 7, and 8, while the crosslinked enzyme retained more than 70% of its initial activity at all pH values after 90 days. Both enzymes showed great potential for industrial use in continuous processes. Although the cross-Linked enzyme showed high operational stability, its lactose conversion was low.

Saccharomyces boulardii is a clinically validated probiotic yeast widely used to treat gastrointestinal disorders. Its natural probiotic effects, combined with the capacity of surviving the harsh gastrointestinal conditions, makes it an attractive chassis for therapeutic protein delivery. However, its engineering has relied on a limited set of genetic tools and lacks systematic validation, which has constrained its development as a next generation engineered probiotic. In this study, the compatibility of the EasyClone-MarkerFree vector system with CRISPR-Cas9 editing in S. boulardii was systematically assessed. Eight genomic loci were identified in silico as suitable for integration, and four were used to construct strains with the Aspergillus niger lacA gene integrated at one or multiple genomic loci, encoding an extracellular β-galactosidase capable of hydrolysing lactose into glucose and galactose, an activity of potential therapeutic relevance for lactose intolerance. Engineered strains efficiently hydrolysed lactose, with the strain with the lacA cassette integrated in three genomic loci (Sb-Triple) displaying up to an 11.76-fold increase in specific β-galactosidase activity relative to single-integration strains, without compromising growth. Importantly, Sb-Triple retained the native pH tolerance characteristic of S. boulardii and was able to grow on, and hydrolyse lactose from, cheese whey powder, a food-grade substrate. These results demonstrate a validated CRISPR-Cas9 strategy for high extracellular β-galactosidase secretion in the clinically established S. boulardii CNCM I-745, while evaluating the compatibility of EasyClone-MarkerFree vector set with this strain. Overall, this work provides a foundation for future studies where the potential of the developed strains is evaluated in clinically relevant settings.

Background: Dairy products provide protein, calcium, and other essential nutrients, but intake may vary across sociodemographic and lifestyle groups. Objective: To characterize dairy consumption patterns and identify factors associated with consuming ≥3 dairy servings/day among adults recruited from selected Peruvian cities. Methods: A cross-sectional study was conducted from April to June 2026. Of the 1619 eligible participants, 203 (12.5%) reported no dairy consumption and were analyzed separately for reasons for non-consumption; the final analytical sample included 1265 dairy consumers. Dairy intake was assessed using a culturally adapted questionnaire with content validity evaluated by expert judgment. Descriptive analyses, bivariate tests, and multivariable logistic regression were performed. Results: Mean age was 33.0 ± 13.0 years, and 61.2% were women. The most commonly consumed products were cheese (90.9%), yogurt (85.4%), whole milk (71.2%), and quesillo or fresh cheese (60.8%). Overall, 16.3% consumed ≥3 dairy servings/day. Participants recruited from cities in the Highlands (aOR = 0.54; 95% CI: 0.37-0.79) and Amazon (aOR = 0.59; 95% CI: 0.40-0.87) had lower odds of reaching this threshold than those from the Coast. Middle socioeconomic status was associated with lower odds (aOR = 0.48; 95% CI: 0.34-0.67), whereas physical activity ≥3 times/week was associated with higher odds (aOR = 1.53; 95% CI: 1.09-2.14). Perceived lactose intolerance was the most frequently reported reason for non-consumption (46.8%). Conclusions: Consumption of ≥3 dairy servings/day was uncommon. Socioeconomic differences were the most consistent correlate, while geographic and physical activity associations warrant cautious interpretation.

Background: Lactose intolerance is a common gastrointestinal condition with variable clinical expression, often requiring both clinical and genetic evaluation. While genetic testing identifies genetic predisposition to lactase persistence or non-persistence, the relationship between specific LCT genotypes and clinical response to dietary intervention remains incompletely understood. Aim: To assess the association between LCT genotypes (CC, CT, TT) and symptom improvement following a lactose-free diet in an adult population and to explore the implications for clinical assessment. Methods: A retrospective observational study was conducted including 538 adult patients evaluated for suspected lactose intolerance. Genotype distribution and its association with clinical response to a lactose-free diet were analyzed. Group comparisons were performed using chi-square and non-parametric tests, and logistic regression analysis was used to identify genotype-specific predictors of symptom improvement. Results: The CC genotype was the most prevalent (70.8%), followed by CT (22.9%) and TT (6.3%). No significant differences were observed between genotype groups regarding age, sex, or area of residence. A significant association was identified between LCT genotype and clinical response to lactose restriction (p < 0.001). CT carriers demonstrated the highest rate of symptom improvement (70.7%), followed by CC (43.8%) and TT (17.6%). Logistic regression analysis confirmed higher odds of improvement among CT carriers (OR = 11.28, 95% CI: 4.30-29.56, p < 0.001) and CC carriers (OR = 3.64, 95% CI: 1.47-9.00, p = 0.005) compared with TT individuals. Conclusions: LCT genotype was significantly associated with clinical response to a lactose-free diet. These findings suggest that genetic variation may contribute to differences in symptom response to dietary lactose restriction and support the use of genotypic information as a complementary component of clinical assessment. However, the observed variability in clinical outcomes across genotype groups highlights the multifactorial nature of lactose intolerance and indicates that genetic predisposition alone does not fully explain symptom expression or dietary response.

This study investigated the potential of hazelnut beverage, obtained from hazelnut cake through high-pressure homogenization (100 MPa), for use in water kefir production. The effects of different sugar concentrations (2.5%, 5%, and 7.5%) on product properties and hazelnut allergenicity were also examined. Sugar addition significantly stimulated microbial growth, resulting in significant increases in Lactobacilli, Lactococci, acetic acid bacteria, and yeast counts compared to the control sample. At the end of fermentation, the pH values of all samples (4.32-4.54) were found to be below the food safety critical limit of pH 4.6. The best water-holding capacity and physical stability were observed in the sample containing 5% sugar. Rheological analyses showed that the samples exhibited pseudoplastic behavior and that sugar addition increased viscosity. The addition of sugar contributed to an increase in the total phenolic content (11.26-15.15 mg GAE/100 mL) and antioxidant activity (DPPH: 16.77%-22.69%; ABTS: 42.68%-46.79%) at the end of fermentation. In the volatile aroma profile evaluation, 47 volatile compounds were tentatively identified based on mass spectral library matching, and increasing sugar concentration resulted in higher relative abundances of ethanol (6.72%-15.51%) and acetic acid (6.29%-13.88%). Allergen levels decreased from 16.6 µg/mL to 8.42-12.97 µg/mL with the addition of sugar. In conclusion, hazelnut cake, a byproduct with low added value, has been transformed into a new, functional, plant-based, low allergenic, and consumer-approved functional beverage. The developed hazelnut-based water kefir offers a significant alternative for individuals with milk protein allergy, lactose intolerance, and vegan/vegetarian consumers. PRACTICAL APPLICATIONS: Hazelnut cake is a low-value by-product from cold-pressed hazelnut oil production. It can be transformed into a better functional beverage using water kefir fermentation. The hazelnut-based water kefir offers a sustainable and eco-friendly way to make use of food industry by-products. It also meets the growing demand for plant-based fermented drinks. Fermentation enhances the beverage's bioactive properties and lowers hazelnut allergen levels, increasing its potential for creating products with better functionality. In addition, the product serves as a great option for vegan and vegetarian consumers, as well as for those with lactose intolerance or milk protein allergies. The findings of this study may aid in creating new non-dairy fermented drinks and help diversify sustainable food products in the beverage industry. Using 5% sugar supplementation could be particularly beneficial for industrial uses because of its positive effects on physical stability, water-holding capacity, and overall product quality.

Oral enzyme replacement therapy for lactase deficiency is limited by the rapid degradation of free β-galactosidase in the gastrointestinal tract. To overcome this, β-galactosidase was encapsulated into polyelectrolyte microcapsules (PMCs) via layer-by-layer assembly using MnCO3 sacrificial templates, with CaCO3-based PMCs as a reference. MnCO3-PMCs achieved 99.4% encapsulation efficiency and retained 85.8% of initial activity, significantly outperforming CaCO3-PMCs (86.0% and 29.7%). Under simulated gastric conditions (pH 2.0, pepsin), the free enzyme and CaCO3-PMCs were completely inactivated, whereas MnCO3-PMCs preserved ~86% activity. In simulated intestinal fluid, MnCO3-PMCs exhibited a 4.3-fold activity increase within the first hour and maintained a 2.5-fold enhancement after 70 h, while the free enzyme progressively inactivated. Furthermore, MnCO3-PMCs demonstrated superior storage stability, retaining 64% of initial activity after 90 days at 4 °C, compared with 25% for CaCO3-PMCs. Although immobilization increased the Michaelis constant, the shift was smaller for MnCO3-PMCs (8.9-fold) than for CaCO3-PMCs (15.2-fold). In conclusion, MnCO3-templated PMCs effectively protect β-galactosidase from gastrointestinal degradation, sustain prolonged catalytic activity, and offer excellent storage stability, highlighting their strong potential for improving oral enzyme replacement therapy in lactose intolerance.