Leveraging the potential of bacterial lateral gene transfer in boosting the efficacy of an edible probiotic prototype yogurt vaccine for COVID-19.

Administration of coronavirus disease-2019 (COVID-19) vaccines with appropriate booster doses through painful injections under clinical supervision was challenging during the recent COVID-19 pandemic. As an alternative solution, we designed a safer, edible probiotic yogurt vaccine prototype (YoVac) that can be orally consumed by circumventing painful injections and clinical supervision. We hypothesized that YoVac prepared using Lactobacillus carrying an antigen coding gene (donor) can transfer the same to other bacteria (recipients) in the human gut microbiome (hgMb) through lateral gene transfer (LGT) for boosted antigen levels potentially triggering a robust immune response. In this study we confirmed the in vitro LGT efficiency of a plasmid (pRBD-Ampr) containing severe acute respiratory syndrome coronavirus-2 (SARS CoV-2) spike protein-receptor binding domain (RBD) coding gene along with an ampicillin-resistance gene (selection marker) from the probiotic Lactobacillus (donor) cultured from homemade yogurt to E. coli and Helicobacter pylori (recipients). Both the donor and recipient bacteria not only exhibited ampicillin-resistance from pRBD-Ampr but also expressed RBD protein. Furthermore, Lactobacillus isolated from YoVac consistently showed the expression of RBD protein over a period of one month confirming the shelf life of our prototype stored at 4 °C. Taken together, our in vitro results provide a preliminary basis for the potential in vivo transfer of RBD coding gene from YoVac to other bacterial species in the hgMb through LGT and may potentially boost the vaccine dosage by delegating them.

Deciphering the metabolism of Lactobacillus delbrueckii subsp. delbrueckii during soy juice fermentation using phenotypic and transcriptional analysis.

In the context of sustainable diet, the development of soy-based yogurt fermented with lactic acid bacteria is an attractive alternative to dairy yogurts. To decipher the metabolism of Lactobacillus delbrueckii subsp. delbrueckii during soy juice (SJ) fermentation, the whole genome of the strain CIRM-BIA865 (Ld865) was sequenced and annotated. Then Ld865 was used to ferment SJ. Samples were analyzed throughout fermentation for their cell number, carbohydrate, organic acid, free amino acid, and volatile compound contents. Despite acidification, the number of Ld865 cells did not rise, and microscopic observations revealed the elongation of cells from 3.6 µm (inoculation) to 36.9 µm (end of fermentation). This elongation was observed in SJ but not in laboratory-rich medium MRS. Using transcriptomic analysis, we showed that the biosynthesis genes of peptidoglycan and membrane lipids were stably expressed, in line with the cell elongation observed, whereas no genes implicated in cell division were upregulated. Among the main sugars available in SJ (sucrose, raffinose, and stachyose), Ld865 only used sucrose. The transcriptomic analysis showed that Ld865 implemented the two transport systems that it contains to import sucrose: a PTS system and an ABC transporter. To fulfill its nitrogen needs, Ld865 probably first consumed the free amino acids of the SJ and then implemented different oligopeptide transporters and proteolytic/peptidase enzymes. In conclusion, this study showed that Ld865 enables fast acidification of SJ, despite the absence of cell division, leads to a product rich in free amino acids, and also leads to the production of aromatic compounds of interest. IMPORTANCE: To reduce the environmental and health concerns related to food, an alternative diet is recommended, containing 50% of plant-based proteins. Soy juice, which is protein rich, is a relevant alternative to animal milk, for the production of yogurt-like products. However, soy "beany" and "green" off-flavors limit the consumption of such products. The lactic acid bacteria (LAB) used for fermentation can help to improve the organoleptic properties of soy products. But metabolic data concerning LAB adapted to soy juice are lacking. The aim of this study was, thus, to decipher the metabolism of Lactobacillus delbrueckii subsp. delbrueckii during fermentation of a soy juice, based on a multidisciplinary approach. This result will contribute to give tracks for a relevant selection of starter. Indeed, the improvement of the organoleptic properties of these types of products could help to promote plant-based proteins in our diet.

Occurrence, antimicrobial susceptibility, and resistance genes of Staphylococcus aureus in milk and milk products in the Arsi highlands of Ethiopia.

BACKGROUND: In Ethiopia, milk production and handling practices often lack proper hygiene measures, leading to the potential contamination of milk and milk products with Staphylococcus aureus (S. aureus), including methicillin-resistant strains, posing significant public health concerns. This study aimed to investigate the occurrence, antimicrobial susceptibility profiles and presence of resistance genes in S. aureus strains isolated from milk and milk products. METHODS: A cross-sectional study was conducted in the Arsi highlands, Oromia, Ethiopia from March 2022 to February 2023. A total of 503 milk and milk product samples were collected, comprising 259 raw milk, 219 cottage cheese, and 25 traditional yogurt samples. S. aureus isolation and coagulase-positive staphylococci enumeration were performed using Baird-Parker agar supplemented with tellurite and egg yolk. S. aureus was further characterized based on colony morphology, Gram stain, mannitol fermentation, catalase test, and coagulase test. Phenotypic antimicrobial resistance was assessed using the Kirby-Bauer disc diffusion method, while the polymerase chain reaction (PCR) was employed for confirming the presence of S. aureus and detecting antimicrobial resistance genes. RESULTS: S. aureus was detected in 24.9% of the milk and milk products, with the highest occurrence in raw milk (40.9%), followed by yogurt (20%), and cottage cheese (6.4%). The geometric mean for coagulase-positive staphylococci counts in raw milk, yogurt, and cottage cheese was 4.6, 3.8, and 3.2 log10 CFU/mL, respectively. Antimicrobial resistance analysis revealed high levels of resistance to ampicillin (89.7%) and penicillin G (87.2%), with 71.8% of the isolates demonstrating multidrug resistance. Of the 16 S. aureus isolates analyzed using PCR, all were found to carry the nuc gene, with the mecA and blaZ genes detected in 50% of these isolates each. CONCLUSION: This study revealed the widespread distribution of S. aureus in milk and milk products in the Arsi highlands of Ethiopia. The isolates displayed high resistance to ampicillin and penicillin, with a concerning level of multidrug resistance. The detection of the mecA and blaZ genes in selected isolates is of particular concern, highlighting a potential public health hazard and posing a challenge to effective antimicrobial treatment. These findings highlight the urgent need to enhance hygiene standards in milk and milk product handling and promote the rational use of antimicrobial drugs. Provision of adequate training for all individuals involved in the dairy sector can help minimize contamination. These measures are crucial in addressing the threats posed by S. aureus, including methicillin-resistant strains, and ensuring the safety of milk and its products for consumers.

Production of set-type probiotic and prebiotic yogurt-like products using Enterococcus faecium and Enterococcus faecalis strains in combination with pumpkin waste.

This study investigated the effect of pumpkin powder (2 %, 4 %, and 6 %) and Enterococcus faecium and Enterococcus faecalis probiotics on the physicochemical, microbiological, and sensory properties of yogurt samples during 28 days of storage at 4 °C. The prebiotic effect of pumpkin powder (Cucurbita pepo) and the probiotic effect of Enterococcus faecium and E. faecalis were determined. Adding pumpkin powder to yogurt did not significantly alter the pH, acidity, fat, protein, and ash content (p > 0.05). Water holding was not changed during the storage time in the samples of probiotic yogurts, but as the pumpkin powder content increased, the water holding capacity also increased (p < 0.05). This situation did lead to a reduction in syneresis (p < 0.05). The lowest gumminess value at the end of storage was found in the D2 sample (p < 0.05), and the highest adhesiveness value was found in the D4 sample (p < 0.05). Furthermore, throughout the 28-day storage period, E. faecium and E. faecalis maintained a live cell count of ≥6 log CFU g-1 in the probiotic product. As a result of the statistical evaluation, there was a decrease in E. faecium in the D4, S2, and S4 samples, and then it increased again (p > 0.05) during the storage time. As a result of the statistical evaluation, it was determined that the smell, consistency in the spoon, consistency in the mouth, flavor, and acidity changes during the storage were not substantial (p > 0.05). In conclusion, it was found that pumpkin, a byproduct of the pumpkin seed industry, has the potential to act as a prebiotic and improve the properties of dairy products. Additionally, the study suggests that E. faecium and E. faecalis strains could be suitable for probiotic yogurts.

Honey Varietals Differentially Impact Bifidobacterium animalis ssp. lactis Survivability in Yogurt through Simulated In Vitro Digestion.

BACKGROUND: Bifidobacterium animalis ssp. lactis DN-173 010/CNCM I-2494 (B. animalis) is a probiotic strain commonly added to yogurt. Yogurt and honey are a popular culinary pairing. Honey improves bifidobacteria survival in vitro. However, probiotic survival in yogurt with honey during in vitro digestion has not been investigated. OBJECTIVES: The study aimed to evaluate the effects of different honey varietals and concentrations on B. animalis survivability in yogurt through in vitro digestion. METHODS: Yogurt with honey or control-treated samples underwent in vitro simulated oral, gastric, and intestinal digestion. B. animalis cells were enumerated on de Man Rogosa and Sharpe (MRS) medium followed by an overlay with a modified selective MRS medium; all underwent anaerobic incubation. B. animalis were enumerated predigestion and after oral, gastric, and intestinal digestion. There were 2 study phases: Phase 1 tested 4 honey varietals at 20% wt/wt per 170 g yogurt, and Phase 2 tested 7 dosages of clover honey (20, 14, 10, 9, 8, 6, and 4% wt/wt) per 170 g yogurt. RESULTS: Similar B. animalis counts were observed between all treatments after oral and gastric digestion (<1 Log colony forming units (CFU)/g probiotic reduction). Higher B. animalis survivability was observed in yogurt with clover honey after exposure to simulated intestinal fluids (∼3.5 Log CFU/g reduction; P < 0.05) compared to all control treatments (∼5.5 Log CFU/g reduction; P < 0.05). Yogurt with 10-20% wt/wt clover honey increased B. animalis survivability after simulated in vitro digestion (≤ ∼4.7 Log CFU/g survival; P < 0.05). CONCLUSIONS: Yogurt with added honey improves probiotic survivability during in vitro digestion. The effective dose of clover honey in yogurt was 10-20% wt/wt per serving (1-2 tablespoons per 170 g yogurt) for increased probiotic survivability during in vitro digestion.

Genetic diversity and phylogenetic relationships of Streptococcus thermophilus isolates from traditional Turkish yogurt: multilocus sequence typing (MLST).

Yogurt, a globally consumed fermented dairy product, is recognized for its taste and potential health benefits attributed to probiotic bacteria, particularly Streptococcus thermophilus. In this study, we employed Multilocus Sequence Typing (MLST) to investigate the genetic diversity and phylogenetic relationships of 13 S. thermophilus isolates from traditional Turkish yogurt samples. We also assessed potential correlations between genetic traits and geographic origins. The isolates were identified as S. thermophilus using VITEK® MALDI-TOF MS, ribotyping, and 16S rRNA analysis methods. MLST analysis revealed 13 different sequence types (STs), with seven new STs for Turkey. The most prevalent STs were ST/83 (n = 3), ST/135 (n = 2), and ST/134 (n = 2). eBURST analysis showed that these isolates mainly were singletons (n = 7) defined as sequence types (STs) that cannot be assigned to any group and differ at two or more alleles from every other ST in the sample. This information suggests that the isolates under study were genetically distinct from the other isolates in the dataset, highlighting their unique genetic profiles within the population. Genetic diversity analysis of ten housekeeping genes revealed polymorphism, with some genes showing higher allelic variation than others. Tajima's D values suggested that selection pressures differed among these genes, with some being more conserved, likely due to their vital functions. Phylogenetic analysis revealed distinct genetic diversity between Turkish isolates and European and Asian counterparts. These findings demonstrate the genetic diversity of S. thermophilus isolates in Turkish yogurt and highlight their unique evolutionary patterns. This research contributes to our understanding of local microbial diversity associated with yogurt production in Turkey and holds the potential for identifyic strains with enhanced functional attributes.

Compatibility of pulse protein in the formulation of plant based yogurt: a review of nutri-functional properties and processing impact.

With the increased environmental concerns and health awareness among consumers, there has been a notable interest in plant-based dairy alternatives. The plant-based yogurt market has experienced rapid expansion in recent years. Due to challenges related to cultivation, higher cost of production and lower protein content researchers have explored the viability of pulse-based yogurt which has arisen as an economically and nutritionally abundant solution. This review aims to examine the feasibility of utilizing pulse protein for yogurt production. The nutritional, antinutritional, and functional characteristics of various pulses were discussed in detail, alongside the modifications in these properties during the various stages of yogurt manufacturing. The review also sheds light on pivotal findings from existing literature and outlines challenges associated with the production of pulse-based yogurt. Pulses have emerged as promising base materials for yogurt manufacturing due to their favorable nutritional and functional characteristics. Further, the fermentation process can effectively reduce antinutritional components and enhance digestibility. Nonetheless, variations in sensorial and rheological properties were noted when different types of pulses were employed. This issue can be addressed by employing suitable combinations to achieve the desired properties in pulse-based yogurt.

Comparison of the impact of saturated fat from full-fat yogurt or low-fat yogurt and butter on cardiometabolic factors: a randomized cross-over trial.

PURPOSE: Dairy foods are often a major contributor to dietary saturated fatty acids (SFA) intake. However, different SFA-rich foods may not have the same effects on cardiovascular risk factors. We compared full-fat yogurt with low-fat yogurt and butter for their effects on cardiometabolic risk factors in healthy individuals. METHODS: Randomized, two-period crossover trial conducted from October 2022 to April 2023 among 30 healthy men and women (15 to receive full-fat yogurt first, and 15 to receive low-fat yogurt and butter first). Participants consumed a diet with 1.5-2 servings of full-fat (4%) yogurt or low-fat (< 1.5) yogurt and 10-15 g of butter per day for 4 weeks, with 4 weeks wash-out when they consumed 1.5-2 servings of low-fat milk. At baseline, and the end of each 4 weeks, fasting blood samples were drawn and plasma lipids, glycemic and inflammatory markers as well as expression of some genes in the blood buffy coats fraction were determined. RESULTS: All 30 participants completed the two periods of the study. Apolipoprotein B was higher for the low-fat yogurt and butter [changes from baseline, + 10.06 (95%CI 4.64 to 15.47)] compared with the full-fat yogurt [-4.27 (95%CI, -11.78 to 3.23)] and the difference between two treatment periods was statistically significant (p = 0.004). Non-high-density lipoprotein increased for the low-fat yogurt and butter [change, + 5.06 (95%CI (-1.56 to 11.69) compared with the full-fat yogurt [change, - 4.90 (95%CI, -11.61 to 1.81), with no significant difference between two periods (p = 0.056). There were no between-period differences in other plasma lipid, insulin, and inflammatory biomarkers or leukocyte gene expression of ATP-binding cassette transporter 1 and CD36. CONCLUSION: This study suggests that short-term intake of SFAs from full-fat yogurt compared to intake from butter and low-fat yogurt has fewer adverse effects on plasma lipid profile. CLINICALTRIALS: GOV: NCT05589350, 10/15/2022.

Soymilk yogurt fermented using Pediococcus pentosaceus TOKAI 759 m improves mice gut microbiota and reduces pro-inflammatory cytokine production.

This study aimed to determine the anti-inflammatory activities and bioactive compounds of soymilk yogurt prepared using Lactiplantibacillus plantarum TOKAI 17 or Pediococcus pentosaceus TOKAI 759 m. Mice were divided into five groups: normal diet (ND), soymilk, soymilk yogurt using L. plantarum TOKAI 17 (SY 17) or P. pentosaceus TOKAI 759 m (SY 759 m), and 0.5 × 109 cells of each starter strain (BC 17 or BC759m). In the SY 759 m group, the serum pro-inflammatory cytokine levels and the cytotoxicity of natural killer cells were attenuated compared to the ND group. In the cecum microbiota, the abundances of butyrate-producing bacteria increased in the SY 759 m and BC 17 groups. Furthermore, SY 759 m metabolites contained high levels of aglycone isoflavone, adenine and showed a significant decrease in CCL-2 and interleukin-6 production in lipopolysaccharide-induced macrophage. In conclusion, soymilk yogurt produced using P. pentosaceus TOKAI 759 m modulates the gut microbiota and can potentially prevent pro-inflammatory cytokine production.

Phosphorus Content of Several Plant-Based Yogurts.

OBJECTIVE: In people with chronic kidney disease (CKD), hyperphosphatemia is a risk factor for mortality. Though unproven, dietary phosphorus control is considered essential in CKD. Although dietary and serum phosphorus are correlated, phosphorus from plant foods rich in phytate is less bioavailable than from animal and processed foods. Yogurt, valued for its low phosphorus and high protein, may be detrimental in CKD due to animal protein content. Plant-based yogurts (PBYs) might offer similar benefits without the downsides of animal protein, but little is known about their phosphorus content. DESIGN AND METHODS: Protein contents and phosphorus additives were gathered from nutrition labels of several PBYs, including almond, cashew, oat, coconut, and soy substrates. Phosphorus content was measured via emission spectrometry by Eurofins (Madison, WI), and the phosphorus-to-protein ratio (PPR) was calculated for each PBY. RESULTS: Phosphorus content was highest in Silk Soy Strawberry, Silk Almond Strawberry, and Siggi's Coconut Mixed Berries, while it was lowest in So Delicious Coconut Strawberry, Oatly Oat Strawberry, Forager Cashew Strawberry, and Kite Hill Almond Strawberry. Ingredient labels revealed that Silk Soy Strawberry, Silk Almond Strawberry, and Oatly Oat Strawberry contained phosphorus additives, and Siggi's Coconut Mixed Berries contained pea protein additives. Though from the same substrate class, So Delicious Coconut Strawberry and Siggi's Coconut Mixed Berries showed significant differences in phosphorus and protein contents. All seven PBYs had higher PPR ratios than dairy yogurts like Stonyfield Organic Oikos Strawberry, Chobani Nonfat Strawberry, and Yoplait Greek Strawberry. CONCLUSION: Low-PPR foods are important for CKD patients. Siggi's Coconut Mixed Berries had the lowest PPR, making it potentially the most desirable for CKD patients. However, there is high variability in PPR among PBYs with the same substrate; therefore, Delicious Coconut Strawberry had the highest PPR, highlighting the importance of product selection for patients with CKD.

A comparison of the quality of plain yogurt and its analog made from coconut flesh extract.

The aim of this study was to compare the quality of plain yogurt made from cow milk (n = 10) and its plant-based analog made from coconut flesh extract (n = 14). Coconut yogurt alternatives were divided into 2 experimental groups based on differences in their color, which were noted after the packages had been opened. The first group included products with a typical white color (n = 8), and the second group comprised products with a grayish pink color (n = 6) that developed as a result of oxidative processes. In comparison with its plant-based analog, plain yogurt was characterized by higher values of lightness (L*), yellowness (b*) and chroma (C*), higher titratable acidity, a higher content of retinol and α-tocopherol, higher nutritional value of fat, and lower values of water-holding capacity (WHC) and redness (a*). Plain yogurt had lower volatile acidity than its plant-based analog with a grayish pink color. A comparison of yogurt analogs with different colors revealed that the product with a grayish pink color was characterized by a lower value of L*, and higher values of a*, b*, C*, and pH. An analysis of its fatty acid profile demonstrated that it also had a higher proportion of C14:0 and C18:1 cis-9; higher total monounsaturated fatty acids content; a lower proportion of C10:0, C12:0, and C18:2; a lower total content of polyunsaturated fatty acids (PUFA) and essential fatty acids; and a lower ratio of PUFA to saturated fatty acids. The yogurt analog with a grayish pink color had a lower total content of tocopherol isoforms than the remaining products. The yogurt analog with a white color had the highest WHC and γ-tocopherol content. Consumers should be aware of the fact that coconut yogurt alternatives may have nonstandard quality attributes. The differences between such products and yogurt made from cow milk should be explicitly communicated to consumers so that they could make informed purchasing decisions.

Metagenomics reveals differences in the composition of bacterial antimicrobial resistance and antibiotic resistance genes in pasteurized yogurt and probiotic bacteria yogurt from China.

Antimicrobial resistance has become a global public health concern, and antibiotic resistance genes (ARG) in food are a research focus. In China, probiotics and pasteurized yogurts are the 2 main types of commercially available yogurt, but the distribution and differences of antibiotic-resistant bacteria and gene types in these products are not well known. This study used a shotgun metagenomic approach to analyze 22 different types of yogurt collected from 9 main yogurt-producing areas in China; each type of yogurt included 8 different batches of samples. The abundance and diversity of bacteria identified in probiotic yogurt were significantly higher than those in pasteurized yogurt, with Acetobacter, Raoultella, and Burkholderia identified as unique and highly abundant genera in probiotic yogurt. Similarly, the abundance of Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa, and Enterobacter spp. was higher than that in pasteurized yogurt. A total of 1,149 ARG subtypes belonging to 16 ARG types were identified, with the highest abundance of rifampicin, multidrug efflux pumps, and quinolone resistance genes detected. Network analysis revealed significant nonrandom co-occurrence relationships between different types and subtypes of ARG in yogurt samples. A total of 44 ARG subtypes in pasteurized yogurt were potentially hosted by 36 bacterial genera, and in probiotic yogurt, 63 ARG were expected to be hosted by 86 bacterial species from 37 genera. These findings indicate potential safety issues in fermented dairy products and emphasize the need for a more hygienic environment when processing probiotic yogurt.

Consumer Perception of Sweeteners in Yogurt.

Consumers are interested in products with reduced sugar, a trend which has been attributed to consumers associating high-sugar foods with obesity, diabetes, heart disease and high blood pressure. The objective of this study was to evaluate consumer perception of sugar reduction in yogurt - both conceptually and for liking of actual products. An online survey (n = 1290) was conducted to evaluate consumer perception of sweeteners available in commercial yogurts. Participants who purchased yogurt at least once in the past 3 mo completed a series of exercises that involved demographics, agreement, Kano, and Maximum Difference (MaxDiff) scaling questions. Subsequently, vanilla 1% milkfat yogurts sweetened with sucrose were formulated to iso-sweet taste intensity with either stevia, allulose, a blend of sucrose and honey or a blend of sucrose, stevia and allulose using magnitude estimate scaling followed by paired comparison tests with consumers (n = 40). A consumer acceptance test (n = 229) was then conducted using the 5 vanilla yogurts. Half of the consumers (n = 115) were primed with the specific yogurt sweetener and the other half were not. Survey and acceptance test data were evaluated by univariate and multivariate analyses. Conceptually, consumers preferred the claim "naturally sweetened" over a reduced sugar claim in yogurt (P < 0.05). Honey was preferred over other sweeteners, followed by agave nectar and cane sugar (P < 0.05). Allulose was the least preferred sweetener option conceptually (P < 0.05). Priming with sweetener type positively impacted consumer acceptance, regardless of sweetener (P < 0.05). When yogurts were tasted, yogurts sweetened with sucrose were the most preferred regardless of priming (P < 0.05). Yogurts sweetened with allulose or a blend of allulose, stevia, and sucrose received higher liking scores than yogurts with stevia (P < 0.05), suggesting that allulose can assist with sugar reduction in yogurt. Consumers are interested in healthy yogurts that are naturally sweetened, but flavor/taste remains the driving force for liking and purchase. The use of a natural non-nutritive sweetener that delivers the sensory experience of sucrose is ultimately more important than familiarity with the actual sweetener.

Engineering the ß-galactosidase from Aspergillus oryzae for making lactose-free and no-sugar-added yogurt.

Yogurt usually contains 5% to 7% sugar and 3% to 5% lactose. As ß-galactosidases can hydrolyze lactose and improve sweetness, they have the potential to produce lactose-free (LF) and no-sugar-added (NSA) yogurt. In this study, the ß-galactosidase AoBgal35A from Aspergillus oryzae was engineered by site-saturation mutagenesis. Results of 19 variants of T955 residue showed that the lactose hydrolysis rate of T955R-AoBgal35A was up to 90.7%, which is much higher than the 78.5% of the wild type. Moreover, the optimal pH of T955R-AoBgal35A was shifted from pH 4.5 to pH 5.5, and the optimal temperature decreased from 60°C to 50°C. The mutant T955R-AoBgal35A was successfully expressed in Komagataella pastoris, which produced extracellularly 4,528 U/mL of ß-galactosidase activity. The mutant T955R-AoBgal35A was used to produce LF yogurt. The Streptococcus thermophilus count of LF yogurt increased from 7.9 to 9.5 log cfu/g, which is significantly higher than that of the control group (8.9 log cfu/g). The residual lactose content of LF yogurt was 0.13%, meeting the requirements of the national standard in China for the "lactose-free" label (<0.5%). Furthermore, sugar in yogurt was replaced by whey powder to produce LF-NSA yogurt. The optimal addition content of whey powder was 7.5%. The texture, water-holding capacity, and titratable acidity of LF and LF-NSA yogurt achieved good shelf life stability. Therefore, this study provides an insight for technological implications of ß-galactosidases in the dairy industry.

Screening of a potential probiotic Lactiplantibacillus plantarum NUC08 and its synergistic effects with yogurt starter.

This study aims to identify lactic acid bacteria (LAB) isolates possessing physiological characteristics suitable for use as probiotics in yogurt fermentation. Following acid and bile salt tolerance tests, Lactiplantibacillus plantarum (NUC08 and NUC101), Lacticaseibacillus rhamnosus (NUC55 and NUC201), and Lacticaseibacillus paracasei (NUC159, NUC216, and NUC351) were shortlisted based on intraspecies distribution for further evaluation. Their physiological probiotic properties, including transit tolerance, adhesion, autoaggregation, surface hydrophobicity, biofilm formation, and antibacterial activity, were assessed. Principal component analysis indicated that Lactiplantibacillus plantarum NUC08 was the preferred choice among the evaluated strains. Subsequent investigations revealed that co-culturing Lactiplantibacillus plantarum NUC08 with 2 yogurt starter strains resulted in a cooperative and synergistic effect, enhancing the growth of mixed strains and increasing their tolerance to simulated gastric and intestinal conditions. Additionally, when Vibrio harveyi bioluminescent reporter strain was used, the 3 cocultured strains cooperated to induce the activity of a quorum sensing (QS) molecule autoinducer-2 (AI-2), hinting a potential connection between phenotypic traits and QS in the cocultured strains. Importantly, LAB viable counts were significantly higher in yogurt co-fermented with Lactiplantibacillus plantarum NUC08, consistently throughout the storage period. In conclusion, the study demonstrates that the probiotic strain Lactiplantibacillus plantarum NUC08 can be employed in synergy with yogurt starter strains, affirming its potential for use in the development of functional fermented dairy products.

Development of probiotic yogurt products incorporated with Lactobacillus kefiranofaciens OSU-BSGOA1 in mono- and coculture with Kluyveromyces marxianus.

The bacterium Lactobacillus kefiranofaciens OSU-BDGOA1 and yeast Kluyveromyces marxianus bdgo-ym6 were previously isolated from kefir grains and have shown probiotic traits in mono- and coculture. This research evaluates the effect of introducing probiotic kefir microorganisms in monoculture and in coculture alongside yogurt starter cultures on the physicochemical and rheological properties, volatile flavor compounds, survival of the microorganisms during simulated digestion, and sensory attributes of the final fermented products. The incorporation of L. kefiranofaciens OSU-BDGOA1 in monoculture showed promising outcomes, resulting in a final product showing more solid-like characteristics and potentially improving the texture of the product. There was also a significant increase in the concentration of desirable volatile flavor compounds in the yogurt with the monoculture, particularly 2,3-butanedione, displaying a positive correlation with buttery flavor in the sensory analysis. The inclusion of L. kefiranofaciens in monoculture also promoted better sensory attributes and was significantly better than the yogurt with the coculture with the yeast, showing promising results for the incorporation of this probiotic bacterium into functional fermented dairy products.

Role and influence mechanism of different concentration of hyaluronic acid on physicochemical and organoleptic properties of yogurt.

Hyaluronic acid (HA) has been approved to be added to milk and other dairy products, it has highly water-binding ability which can combine with a large number of water molecules through intramolecular hydrogen bonding to form high viscous gels. In addition, HA is one of the prebiotics, can provide health benefits like anti-aging, anti-inflammatory, angiogenic, is a potential additive for enhancing the quality of yogurt. Therefore, the aim of this study was to evaluate the effect of 0%, 0.02%, 0.05%, 0.1%, 0.25% and 0.5% HA on rheological properties, functional properties, thermal stability, protein stability, protein structure and protein fractions of yogurt. The addition of HA, storage modulus (G') is always larger than loss modulus (G'') in all the samples, which is a typical characteristic of gel networks, and the microstructures of the yogurt samples showed a continuous and more homogeneous spatial network structure. Overall, the higher concentration (0.5%) had positive effect on the yogurt characteristics, like higher WHC, foam stability, microstructure, and texture. In contrast, the 0.1% concentration HA lead to a very abnormal results, it had a negative effect on yogurt including water-holding capacity, texture, and protein stability, suggesting structural destabilization and disruption of inter-aggregation before protein. These findings provide a valuable fundamental data for commercialized HA adding yogurt development and quality control processes.

Graduate Student Literature Review: Network of flavor compounds formation and influence factors in yogurt.

Yogurt is popular as a natural and healthy food, but its flavor greatly affects acceptability by consumers. Flavor compounds of yogurt is generally produced by the metabolism of lactose, protein and fat, and the resulting flavors include carbonyls, acids, esters and alcohols, etc. Each flavor compounds could individually provide the corresponding flavor, or it can be combined with other compounds to form a new flavor. The flavor network was formed among the metabolites of milk components, and acetaldehyde, as the central compounds, played a role in connecting the whole network. The flavor compounds can be affected by many factors, such as the use of different raw milks, ways of homogenization, sterilization, fermentation, post ripening, storage condition and packaging materials, etc., which can affect the overall flavor of yogurt. This paper provides an overview of the volatile flavor compounds in yogurt, the pathways of production of the main flavor compounds during yogurt fermentation, and the factors that influence the flavor of yogurt including type of raw milk, processing, and storage. It also tries to provide theoretical guidance for the product of yogurt in ideal flavor, but further research is needed to provide a more comprehensive description of the flavor system of yogurt.

Optimizing Skim Milk Yogurt Properties: Combined Impact of Trans-glutaminase and Protein-Glutaminase.

The lack of fat in yogurt can lead to alterations in taste and whey separation, reducing consumer acceptance. In this study, the feasibility of enhancing the quality of skim milk yogurt through a combination of transglutaminase (TG) and protein-glutaminase (PG) was investigated. The combination of TG and PG resulted in simultaneous cross-linking and deamidated of casein micelles, with PG deamidation taking priority over TG cross-linking, leading to higher solubility and lower turbidity of milk proteins compared with TG alone. When 0.06 U/mL TG and 0.03 U/mL PG were added, firmness and viscosity indexes significantly increased by 38.26 and 78.59%, respectively as compared with the control. Microscopic images revealed increased cross-linking with casein and filling of cavities by smaller sub-micelles in the combination of TG and PG treatment. Furthermore, the combination of TG and PG resolved issues of rough taste and whey separation, leading to improved overall liking. This study highlights the benefits of using both enzymes in dairy production and has important implication for future research.

Comparative transcriptomic analysis of the flavor production mechanism in yogurt by traditional starter strains.

The synergistic fermentation of milk by Streptococcus thermophilus and Lactobacillus delbrueckii ssp. bulgaricus is one of the key factors that determines the quality of yogurt. In this study, the mechanism whereby yogurt flavor compounds are produced by a mixture of S. thermophilus SIT-20.S and L. delbrueckii ssp. bulgaricus SIT-17.B were investigated by examining the flavor production, growth, and gene transcription of these strains. The results showed that yogurt produced by a 10:1 mixture of the aforementioned strains had the highest abundance of acetoin, whereas yogurt produced by a 1:1 mixture had the highest abundance of diacetyl and acetaldehyde. In addition, the growth of S. thermophilus SIT-20.S was enhanced in the 10:1 mixture. Transcriptomic analysis revealed differentially expressed genes in the flavor-compound-related pathways of S. thermophilus SIT-20.S and L. delbrueckii ssp. bulgaricus SIT-17.B in yogurts produced by 10:1 and 1:1 mixtures compared with those produced by either strain alone. Mixed fermentations regulated the expression of genes related to glycolysis, resulting in an increase of pyruvate, which is an important precursor for diacetyl and acetoin synthesis. The gene encoding the acetoin reductase (SIT-20S_orf01454) was decreased in S. thermophilus SIT-20.S, which ensured the accumulation of acetoin. In addition, the gene encoding the acetaldehyde dehydrogenase (SIT-20S_orf00949) was upregulated in S. thermophilus SIT-20.S, and the expression of alcohol dehydrogenase (SIT-20S_orf01479; SIT-17B_orf00943) was downregulated in both strains, maintaining the abundance of acetaldehyde. In addition, the gene encoding the NADH oxidase (SIT-17B_orf00860) in L. delbrueckii ssp. bulgaricus SIT-17.B were upregulated, which promoted the accumulation of diacetyl and acetoin. Overall, we characterized the mechanism by which S. thermophilus and L. delbrueckii ssp. bulgaricus synergistically generated yogurt flavor compounds during their production of yogurt and highlighted the importance of appropriate proportions of fermentation starters for improving the flavor of yogurts.