Carao is considered a functional ingredient since its bioactive compounds are meaningful in nutritional, pharmacological, and medicinal applications. The objective of this study was to determine the effects of carao pulp powder on the bacterial viability, acid tolerance, bile tolerance, and protease activity of S. thermophilus STI-06 and L. bulgaricus LB-12. M17 broth with 0.5% lactose and MRS broth were used for S. thermophilus and L. bulgaricus, respectively, for determining bacterial viability, acid tolerance, and bile tolerance. Skim milk was used to study the protease activity of both bacteria. The carao was added at 0 (control), 1.3, 2.6, and 5.3 (g/L) into the broths and skim milk. The broths were enumerated for bacterial viability (every 2 h), bile tolerance (every 4 h), and acid tolerance (every 30 min), and the skim milk was analyzed for protease activity (every 12 h). The General Linear Model (PROC GLM) was used to analyze the data. The 2.6 g/L and 5.3 g/L usage level of carao improved the acid tolerance of S. thermophilus. Carao did not affect the acid tolerance of L. bulgaricus. The usage of 5.3 g/L of carao significantly improved the bile tolerance and protease activity of both bacteria. However, carao did not affect the viability of either bacteria. Overall, 5.3 g/L of carao with these probiotics could be recommended in fermentation processes.

The impact of yogurts made with starter culture bacteria (L. bulgaricus and S. thermophilus) and supplemented with ingredients (maitake mushrooms, quercetin, L-glutamine, slippery elm bark, licorice root, N-acetyl-D-glucosamine, zinc orotate, and marshmallow root) that can help treat leaky gut were investigated using the Caco-2 cell monolayer as a measure of intestinal barrier dysfunction. Milk from the same source was equally dispersed into nine pails, and the eight ingredients were randomly allocated to the eight pails. The control had no ingredients. The Caco-2 cells were treated with isoflavone genistein (negative control) and growth media (positive control). Inflammation was stimulated using an inflammatory cocktail of cytokines (interferon-γ, tumor necrosis factor-α, and interleukin-1β) and lipopolysaccharide. The yogurt without ingredients (control yogurt) was compared to the yogurt treatments (yogurts with ingredients) that help treat leaky gut. Transepithelial electrical resistance (TEER) and paracellular permeability were measured to evaluate the integrity of the Caco-2 monolayer. Transmission electron microscopy (TEM), immunofluorescence microscopy (IM), and real-time quantitative polymerase chain reaction (RTQPCR) were applied to measure the integrity of tight junction proteins. The yogurts were subjected to gastric and intestinal digestion, and TEER was recorded. Ferrous ion chelating activity, ferric reducing potential, and DPPH radical scavenging were also examined to determine the yogurts’ antioxidant capacity. Yogurt with quercetin and marshmallow root improved the antioxidant activity and TEER and had the lowest permeability in fluorescein isothiocyanate (FITC)–dextran and Lucifer yellow flux among the yogurt samples. TEM, IM, and RTQPCR revealed that yogurt enhanced tight junction proteins’ localization and gene expression. Intestinal digestion of the yogurt negatively impacted inflammation-induced Caco-2 barrier dysfunction, while yogurt with quercetin, marshmallow root, maitake mushroom, and licorice root had the highest TEER values compared to the control yogurt. Yogurt fortification with quercetin, marshmallow root, maitake mushroom, and licorice root may improve functionality when dealing with intestinal barrier dysfunction.

Consumers are becoming aware of functional ingredients such as medicinal herbs, polyphenols, mushrooms, amino acids, proteins, and probiotics more than ever before. Like yogurt and its probiotics, L-glutamine, quercetin, slippery elm bark, marshmallow root, N-acetyl-D-glucosamine, licorice root, maitake mushrooms, and zinc orotate have demonstrated health benefits through gut microbiota. The impact of these ingredients on yogurt starter culture bacteria characteristics is not well known. The objective of this study was to determine the influence of these ingredients on the probiotic characteristics, tolerance to gastric juices and lysozyme, protease activity, and viability of Streptococcus thermophilus STI-06 and Lactobacillus bulgaricus LB-12. Acid tolerance was determined at 0, 30, 60, 90, and 120 min of incubation, whereas bile tolerance was analyzed at 0, 4, and 8 h. The microbial growth was determined at 0, 2, 4, 6, 8, 10, 12, 14, and 16 h of incubation, while protease activity was evaluated at 0, 12, and 24 h. The application of marshmallow root, licorice root, and slippery elm bark improved bile tolerance and acid tolerance of S. thermophilus. These ingredients did not impact the bile tolerance, acid tolerance, and simulated gastric juice tolerance characteristics of L. bulgaricus over 8 h and 120 min (respectively) of incubation. Similarly, the growth of S. thermophilus and L. bulgaricus was not affected by any of these functional ingredients. The application of marshmallow root, N-acetyl-D-glucosamine, and maitake mushroom significantly increased the protease activity of S. thermophilus, whereas the protease activity of L. bulgaricus was not affected by any ingredient. Compared to the control, marshmallow root and quercetin samples had higher mean log counts and log counts for S. thermophilus on the simulated gastric juice and lysozyme resistance in vitro test, respectively. For L. bulgaricus, licorice root, quercetin, marshmallow root, and slippery elm bark samples had higher log counts than the control samples.

Frozen yogurts contain yogurt culture bacteria, which might impart health benefits to their consumers. Global frozen yogurt market sales are expected to grow by 4.8% by 2028, which represents an important opportunity for the industry, consumers and researchers. Polyphenols are metabolites found in plants which have antioxidant and anti-inflammatory properties and might prevent chronic diseases such as cancer, diabetes and cardiovascular diseases. The objective of this study was to elucidate the effect of the polyphenol hesperidin on the physico-chemical, microbiological and sensory characteristics of frozen yogurts. Hesperidin was incorporated into frozen yogurt at three concentrations (125, 250 and 500 mg/90 g of product), while yogurt with no hesperidin was used as a control. The viscosity and overrun of the frozen yogurt were analyzed on day 0. The hardness, pH, color and Lactobacillus bulgaricus and Streptococcus thermophilus counts were determined after 0, 30 and 60 d. The melting rate was determined at 60 and 90 min after 0, 30 and 60 d. The bile and acid tolerances of both S. thermophilus and L. bulgaricus were measured after 7 and 60 d. A hedonic scale of nine points was used to measure sensory attributes. Data were analyzed at α = 0.05 with an ANOVA with Tukey’s adjustment, and McNemar’s test was used to analyze purchase intent. Hesperidin did not influence the pH, overrun or microbial characteristics. Polyphenol addition compared to the control decreased the melting rate but increased the hardness and bile tolerance of L. bulgaricus, as well as the L* and b* values. The sensory characteristics were not influenced by the lowest concentration of hesperidin, as it was not statistically different from the control. Moreover, consumers were interested in purchasing frozen yogurt with added hesperidin after learning about the health claim. This study can assist in the development of a healthier frozen yogurt in an increasingly competitive market.

The current research aimed to evaluate the potential effects of Solanum mammosum, Dioon mejiae, and Amanita caesarea on Streptococcus thermophilus and Lactobacillus delbrueckii subsp. bulgaricus survival and performance after exposure to different harsh conditions such as bile, acid, gastric juice, and lysozyme to mimic the digestive system from mouth to the intestine. Probiotic protease activity was observed to evaluate the proteolytic system. Probiotics were cultured in a broth mixed with plant material, and after incubation, the results were compared to the control sample. Therefore, plant material’s total phenolic compound, total carotenoid compound, antioxidant activity, sugar profile, and acid profile were obtained to discuss their impact on the survival of probiotics. The results indicate that Amanita caesarea negatively affected probiotic survival in the bile tolerance test and positively affected Lactobacillus bulgaricus in the protease activity test. Otherwise, the other plant material did not change the results significantly (p > 0.05) compared to the control in different tests. Consequently, Solanum mammosum and Dioon mejiae had no significant effects (p > 0.05) in increasing probiotic survival.

Carao (Cassia grandis) contains numerous bioactive substances that contribute to gastrointestinal well-being. The present study assessed the potential impacts of carao on the viability and performance of Streptococcus thermophilus and Lactobacillus bulgaricus under various adverse conditions. These conditions included bile, acid, gastric juice, and lysozyme exposure, simulating the digestive process from the mouth to the intestines. The activity of proteases from cultures was monitored to examine their proteolytic capabilities. To achieve this, the cultures were cultivated in a solution containing plant material, and the results were compared against a control sample after an incubation period. Subsequently, the total phenolic content, total carotenoid content, antioxidant activity, sugar profile, and acid profile of the plant materials were analyzed. These analyses were conducted to explore these compounds’ influence on cultures’ survival. Seeds contained the highest total phenols (766.87 ± 11.56 µg GAE/mL), total carotenoid content (7.43 ± 0.31 mg Q/mL), and antioxidant activity (40.76 ± 1.87%). Pulp contained the highest moisture (12.55 ± 0.44%), ash (6.45 ± 0.15%), lipid (0.66 ± 0.07%), protein (16.56 ± 0.21%), sucrose (9.07 ± 0.78 g/100 g), and fructose (3.76 ± 0.06 g/ 100 g). The crust had the highest content of ash (85.14 ± 0.27%) and succinic acid (2.01 ± 0.06 g/100 g). Results indicated that seeds negatively affected cultures’ survival in the bile tolerance test and had positive effects on Lactobacillus bulgaricus in the protease activity test. Otherwise, the other carao tissues could not change the results significantly (p > 0.05) compared to the control in different tests. The carao crust positively affected cultures’ against protease activity, especially in Lactobacillus bulgaricus, and had a negative effect on the growth of S. thermophilus in the lysozyme and gastric acid resistance test.