Postbiotic potential of Bacillus velezensis KMU01 cell-free supernatant for the alleviation of obesity in mice.

Attention toward the preventive effects of postbiotics on metabolic diseases has increased because of greater stability and safety over probiotics. However, studies regarding the bioactive effects of postbiotics, especially from probiotic Bacillus strains, are relatively limited. The anti-obesity effects of the cell-free culture supernatant of Bacillus velezensis KMU01 (CFS-B.vele) were evaluated using high-fat-diet (HFD)-induced mice. HFD-induced mice (n = 8 per group) received equal volumes of (1) CFS-B.vele (114 mg/kg) in PBS, (2) Xenical in PBS, or (3) PBS alone by oral gavage daily for 13 weeks. The results demonstrated that CFS-B.vele changed the gut microbiota and showed anti-obesity effects in HFD-induced obese mice. The elevated Firmicutes/Bacteroidota ratio induced by HFD was decreased in the CFS-B.vele group compared to the other groups (p < 0.05). The CFS-B.vele intervention led to the enrichment of SCFA-producers, such as Roseburia and Eubacterium, in the cecum, suggesting their potential involvement in the amelioration of obesity. Due to these changes, the various obesity-related biomarkers (body weight, fat in tissue, white adipose tissue weight and size, serum LDL-cholesterol level, hepatic lipid accumulation, and adipogenesis/lipogenesis-related gene/protein expression) were improved. Our findings suggest that CFS-B.vele has potential as a novel anti-obesity agent through modulation of the gut microbiota.

Synergistic antimicrobial effect and mode of action of palmarosa oil-loaded nanoemulsion and citric acid against Pectobacterium carotovorum.

The synergistic antimicrobial activity of palmarosa oil (Cymbopogon martini, PO)-loaded nanoemulsion (PO-NE) and citric acid (CA) against Pectobacterium, the major pathogen for soft-rot disease, was evaluated. The combination of PO-NE and CA (PO-NE + CA) significantly improved the storage stability of PO-NE at 30 °C. Compared to the anti-Pectobacterium activity of alone, PO-NE + CA reduced the minimum inhibitory concentration (MIC) by 1/4 and 1/2, respectively. Bactericidal efficacy of PO-NE + CA against P. carotovorum PCC3 was similar of PO-NE alone in the MIC in time-kill kinetic assay. PO-NE treatment mainly influenced membrane integrity, while CA treatment strongly stimulated intracellular ATP depletion. This synergistic combination effectively reduced the use of PO-NE, imparting a strong flavor note without sacrificing the antimicrobial efficacy against Pectobacterium. Supplementary Information: The online version contains supplementary material available at 10.1007/s10068-022-01217-6.

Erratum to: Effect of Modified Casein to Whey Protein Ratio on Dispersion Stability, Protein Quality and Body Composition in Rats.

[This corrects the article DOI: 10.5851/kosfa.2021.e42.].

Effects of Lactobacillus reuteri MG5346 on Receptor Activator of Nuclear Factor-Kappa B Ligand (RANKL)-Induced Osteoclastogenesis and Ligature-Induced Experimental Periodontitis Rats.

Effects of culture supernatants of Lactobacillus reuteri MG5346 (CS-MG5346) on receptor activator of nuclear factor-kappa B ligand (RANKL)-induced osteoclastogenesis were examined. CS-MG5346 treatment up to 400 µg/mL significantly reduced tartrate-resistant acid-phosphatase (TRAP) activity, the phenotype biomarker of osteoclast, without affecting cell viability. CS-MG5346 inhibited the expression of osteoclast specific transcriptional factors (c-fos and nuclear factor-activated T cells c1) and their target genes (TRAP, cathepsin, and matrix metallo-proteinase-9) in a dose-dependent manner (p<0.05). The administration of L. reuteri MG5346 (2×108 CFU/day) for 8 wks significantly improved furcation involvement, but no difference was observed in alveolar bone loss in ligature-induced experimental periodontitis rats. The elevated RANKL/ osteoprotegerin ratio, the biomarker of periodontitis, was significantly lowered in the gingival tissue by administration of L. reuteri MG5346 (p<0.05). L. reuteri MG5346 showed excellent stability in simulated stomach and intestinal fluids and did not have antibiotic resistance. Based on the results, L. reuteri MG5346 has the potential to be a promising probiotic strain for oral health.

Effect of phospholipid matrix on emulsion stability, microstructure, proteolysis, and in vitro digestibility in model infant formula emulsion.

The effects of adding different phospholipid (PL) matrices [milk sphingomyelin (SM) vs soy phosphatidylcholine (PC)] on emulsion stability, microstructure, and in vitro simulated lipid digestion were examined using a Model Infant Formula Emulsion (MIFE). The emulsion stability of MIFE increased significantly with PL addition (0.1 and 0.2 %). Compared to sole MIFE or MIFE + PC, the incorporation of SM resulted in increased emulsion stability (p < 0.05) and a greater amount of free fatty acid release (p < 0.05) during in vitro simulated digestion. This was mainly due to the reduction of intensive droplet aggregation, thus providing a large surface area and improved digestibility. This is further experimentally supported by the evolution of particle size distribution, zeta-potential, and microstructure analysis using confocal laser scanning microscopy. The incorporation of SM in the emulsion formation significantly delayed digestion of ß-lactoglobulin during in vitro digestion. Lipid digestibility in MIFE was altered depending on the type of PL matrix, and SM displayed a superior effect to PC. Thus, the creation of a novel emulsion interface by the appropriate selection of emulsifiers can be used to improve lipid digestion in infants and obtain desirable nutritional consequences.

Roles of Milk Fat Globule Membrane on Fat Digestion and Infant Nutrition.

Milk fats are present as globules emulsified in the aqueous phase of milk and stabilized by a delicate membrane architecture called milk fat globule membrane (MFGM). The unique structure and composition of the MFGM play an important role in fat digestion and the metabolic programming of neonates. The objective of this review is to compare the structure, composition, and physicochemical characteristics of fat globules in human milk, bovine milk, and infant formula. It provides an overview of the fat digestion process and enzymes in healthy infants, and describes the possible roles of the MFGM in association with factors affecting fat digestion. Lastly, the health benefits of the MFGM on infant nutrition and future perspectives are discussed with a focus on brain development, metabolic response, and gut health.

The interaction of milk sphingomyelin and proteins on stability and microstructure of dairy emulsions.

The interaction between dairy proteins [micellar casein (MC) vs. whey protein isolate (WPI)] and phospholipids [PL; soy phosphatidylcholine (PC) vs. milk sphingomyelin (SM)] in an oil-in-water emulsion system was investigated. Sole PC-stabilized emulsion (1%, wt/vol) showed a significantly larger mean particle diameter (6.5 µm) than SM-stabilized emulsions (3.8 µm). The mean particle diameters of emulsions prepared by the combination of protein (1%, wt/vol) and PL (1%, wt/vol) did not significantly differ from the emulsions prepared with a single emulsifier (MC, WPI, and SM). Emulsion instability differed significantly among samples by a centrifugation-mediated accelerated stability test. Emulsion instability increased in the order of MC+SM < MC+PC, WPI+SM < WPI+PC < MC < SM < WPI < PC. Protein surface load determined by aqueous phase depletion was significantly decreased only in WPI+PC emulsion, whereas no significant difference was found between the MC+SM and WPI+SM emulsions. Topographic and phase images of emulsion surface by atomic force microscopy showed surface layers prepared by protein+PL combinations were composites with different mechanical properties, and PL formed a more compact domain than proteins. A smoother phase image was observed in MC+PL combinations than in WPI+PL counterparts. Based on the microstructure analysis using confocal laser scanning microscopy, combination and MC+SM formed a uniform and thick surface coating of fat droplets. More PC aggregates were observed in the emulsions containing PC (sole PC, MC+PC, and WPI+PC) compared with their SM counterparts. Based on these results, the appropriate selection of the PL matrix is important to modulate the emulsion stability of dairy emulsion products.

Effects of Lactobacillus curvatus MG5246 on inflammatory markers in Porphyromonas gingivalis lipopolysaccharide-sensitized human gingival fibroblasts and periodontitis rat model.

This study investigated the effects of Lactobacillus curvatus MG5246 on periodontitis inflammation. Cell-free supernatants (CFS) prepared from L. curvatus MG5246 decreased prostaglandin E2 production and cyclooxygenase-2 gene expression by 60% and 78% in Porphyromonas gingivalis-lipopolysaccharide stimulated human gingival fibroblasts at 400 µg/mL. Gene expressions of tumor necrosis factor-α, interleukin-6, matrix metalloproteinases, and chemokines were significantly downregulated by CFS treatment (p < 0.05). L. curvatus MG5246 (2 × 108 CFU/day, 8 weeks) administration significantly improved alveolar bone loss in the ligature-induced periodontitis rat model. Elevated mRNA expression of the receptor activator of nuclear factor-κB ligand/osteoprotegerin ratio in the gingival tissue was significantly decreased by L. curvatus MG5246 administration (p < 0.05). Moreover, L. curvatus MG5246 showed sufficient tolerance in simulated gastrointestinal conditions (gastric tolerance: 89.48%, intestinal tolerance: 98.62%) and did not show antibiotic resistance and hemolytic activity. Therefore, L. curvatus MG5246 has the potential as novel oral probiotics.

Effect of Modified Casein to Whey Protein Ratio on Dispersion Stability, Protein Quality and Body Composition in Rats.

The present study was designed to investigate the effects of protein formula with different casein (C) to whey protein (W) ratios on dispersion stability, protein quality and body composition in rats. Modification of the casein to whey protein (CW) ratio affected the extent of protein aggregation, and heated CW-2:8 showed a significantly increased larger particle (>100 µm) size distribution. The largest protein aggregates were formed by whey protein self-aggregation. There were no significant differences in protein aggregation when the CW ratios changed from 10:0 to 5:5. Based on the protein quality assessment (CW-10:0, CW-8:2, CW-5:5, and CW-2:8) for four weeks, CW-10:0 showed a significantly higher feed intake (p<0.05), but the high proportion of whey protein in the diet (CW-5:5 and CW-2:8) increased the feed efficiency ratio, protein efficiency ratio, and net protein ratio compared to other groups. Similarly, CW-2:8 showed greater true digestibility compared to other groups. No significant differences in fat mass and lean mass analyzed by dual-energy x-ray absorptiometry were observed. A significant difference was found in the bone mineral density between the CW-10:0 and CW-2:8 groups (p<0.05), but no difference was observed among the other groups. Based on the results, CW-5:5 improved protein quality without causing protein instability problems in the dispersion.

Ecklonia cava Extract Exerts Anti-Inflammatory Effect in Human Gingival Fibroblasts and Chronic Periodontitis Animal Model by Suppression of Pro-Inflammatory Cytokines and Chemokines.

Periodontitis is one of the most common chronic inflammatory diseases. The anti-inflammatory effect of the extract from brown algae Ecklonia cava was analyzed in lipopolysaccharide (LPS)-stimulated human gingival fibroblasts (HGF-1), the most abundant cells in gingival tissue. The gene expressions of cyclooxygenase-2 and interleukin-6 were decreased by 78 and 50%, respectively, at 100 µg/mL Ecklonia cava extract (ECE) treatment. The gene expressions of matrix metalloproteases (MMP-2 and MMP-8) and chemokines (macrophage inflammatory protein 1-alpha and stromal cell-derived factor 1) were also significantly down-regulated by ECE treatment (p < 0.05). The increased reactive oxygen species (ROS) production in HGF-1 cells by LPS stimulation was decreased by 30% at 100 µg/mL ECE treatment. The mitogen-activated protein kinase pathway and the nuclear factor-kappa B (NF-κB) signal activated by ROS were suppressed by ECE in a dose-dependent manner. ECE treatment (400 mg/kg, 8 weeks) significantly improved alveolar bone resorption in the ligature-induced chronic periodontitis rat model. ECE supplementation also lowered elevated mRNA expression of the receptor activator of nuclear factor-kappa B (RANKL)/osteoprotegerin (OPG) in the gingival tissue (p < 0.05). Therefore, ECE mitigated gingival tissue destruction and bone resorption associated with chronic periodontitis condition.

Changes in Particle Size, Sedimentation, and Protein Microstructure of Ultra-High-Temperature Skim Milk Considering Plasmin Concentration and Storage Temperature.

Age gelation is a major quality defect in ultra-high-temperature (UHT) pasteurized milk during extended storage. Changes in plasmin (PL)-induced sedimentation were investigated during storage (23 °C and 37 °C, four weeks) of UHT skim milk treated with PL (2.5, 10, and 15 U/L). The increase in particle size and broadening of the particle size distribution of samples during storage were dependent on the PL concentration, storage period, and storage temperature. Sediment analysis indicated that elevated storage temperature accelerated protein sedimentation. The initial PL concentration was positively correlated with the amount of protein sediment in samples stored at 23 °C for four weeks (r = 0.615; p < 0.01), whereas this correlation was negative in samples stored at 37 °C for the same time (r = -0.358; p < 0.01) due to extensive proteolysis. SDS-PAGE revealed that whey proteins remained soluble over storage at 23 °C for four weeks, but they mostly disappeared from the soluble phase of PL-added samples after two weeks' storage at 37 °C. Transmission electron micrographs of PL-containing UHT skim milk during storage at different temperatures supported the trend of sediment analysis well. Based on the Fourier transform infrared spectra of UHT skim milk stored at 23 °C for three weeks, PL-induced particle size enlargement was due to protein aggregation and the formation of intermolecular ß-sheet structures, which contributed to casein destabilization, leading to sediment formation.

Effects of Probiotic Culture Supernatant on Cariogenic Biofilm Formation and RANKL-Induced Osteoclastogenesis in RAW 264.7 Macrophages.

Postbiotics are a promising functional ingredient that can overcome the limitations of viability and storage stability that challenge the production of probiotics. To evaluate the effects of postbiotics on oral health, eight spent culture supernatants (SCSs) of probiotics were prepared, and the effects of SCSs on Streptococcus mutans-induced cariogenic biofilm formation and the receptor activator of the nuclear factor κB ligand (RANKL)-induced osteoclastogenesis were evaluated in RAW 264.7 macrophages. SCS of Lactobacillus salivarius MG4265 reduced S. mutans-induced biofilm formation by 73% and significantly inhibited tartrate-resistant acid phosphatase (TRAP) activity, which is a biomarker of mature osteoclasts in RAW 264.7 macrophages. The suppression of RANKL-induced activation of mitogen activated the protein kinases (c-Jun N-terminal kinase, extracellular signal-regulated kinase, and p38) and nuclear factor κB pathways, as well as the upregulation of heme oxygenase-1 expression. The suppression of RANK-L-induced activation of mitogen also inhibited the expression of transcriptional factors (c-fos and nuclear factor of activated T cells cytoplasmic 1) and, subsequently, osteoclastogenesis-related gene expression (tartrate-resistant acid phosphatase-positive (TRAP), cathepsin K, and matrix metalloproteinase-9).Therefore, SCS of L. salivarius MG4265 has great potential as a multifunctional oral health ingredient that inhibits biofilm formation and suppresses the alveolar bone loss that is associated with periodontitis.

The Effect of Chrysin-Loaded Phytosomes on Insulin Resistance and Blood Sugar Control in Type 2 Diabetic db/db Mice.

Although a variety of beneficial health effects of natural flavonoids, including chrysin, has been suggested, poor solubility and bioavailability limit their practical use. As a promising delivery system, chrysin-loaded phytosomes (CPs) were prepared using egg phospholipid (EPL) at a 1:3 molar ratio and its antidiabetic effects were assessed in db/db diabetic mice. Male C57BLKS/J-db/db mice were fed a normal diet (control), chrysin diet (100 mg chrysin/kg), CP diet (100 mg chrysin equivalent/kg), metformin diet (200 mg/kg) or EPL diet (vehicle, the same amount of EPL used for CP preparation) for 9 weeks. Administration of CP significantly decreased fasting blood glucose and insulin levels in db/db mice compared with the control. An oral glucose tolerance test and homeostatic model assessment for insulin resistance were significantly improved in the CP group (p < 0.05). CP treatment suppressed gluconeogenesis via downregulation of phosphoenolpyruvate carboxykinase while it promoted glucose uptake in the skeletal muscle and liver of db/db mice (p < 0.05). The CP-mediated improved glucose utilization in the muscle was confirmed by upregulation of glucose transporter type 4, hexokinase2 and peroxisome proliferator-activated receptor γ during treatment (p < 0.05). The CP-induced promotion of GLUT4 plasma translocation was confirmed in the skeletal muscle of db/db mice (p < 0.05). Based on the results, CP showed greater antidiabetic performance compared to the control by ameliorating insulin resistance in db/db mice and phytosome can be used as an effective antidiabetic agent.

Effect of Homogenization Pressure on Plasmin Activity and Mechanical Stress-Induced Fat Aggregation of Commercially Sterilized Ultra High Temperature Milk during Storage.

Commercially sterilized ultra high temperature (UHT) milk was manufactured at different homogenization pressures (20, 25, and 30 MPa), and changes in fat particle size, mechanical stress-induced fat aggregation, plasmin activity, and lipid oxidation were monitored during ambient storage of the UHT milk for up to 16 wk. The particle sizes of milk fat globules were significantly decreased as homogenization pressure increased from 20 to 30 MPa (p<0.05). The presence of mechanical stress-induced fat aggregates in milk produced at 20 MPa was significantly higher than for UHT milk produced at either 25 or 30 MPa. This difference was maintained all throughout the storage. There were no significant differences in plasmin activity, trichloroacetic acid (12%, w/v) soluble peptides, and the extent of lipid oxidation. Based on these results, an increase of homogenization pressure from 20 (the typical homogenization pressure employed in the Korea dairy industry) to 25-30 MPa significantly decreased mechanical stress-induced fat aggregation without affecting susceptibility to lipid oxidation during storage.

Characteristics and Glucose Uptake Promoting Effect of Chrysin-Loaded Phytosomes Prepared with Different Phospholipid Matrices.

Chrysin-loaded phytosomes (CP) were prepared using either soya phosphatidylcholine (SPC) or egg phospholipid (EPL) by the solvent evaporation method. Different phospholipid matrices resulted in significant differences in size, mechanical property and solubility of the CP. The most stable CP was obtained with EPL at a molar ratio of 1:3 (chrysin: EPL, CEP-1:3). CEP-1:3 displayed an average size of 117 nm with uniform size distribution (polydispersity index: 0.30) and zeta potential of -31 mV. A significantly greater elastic modulus of CEP-1:3 (2.7-fold) indicated tighter packing and strong molecular bonding than those of CP prepared with SPC (CSP-1:3). X-ray diffraction and Fourier transform infrared spectroscopic analysis of CEP-1:3 confirmed molecular complexation. CEP-1:3 displayed a greater glucose uptake promoting effect than free chrysin and CSP-1:3 in muscle cells by stimulating gene expression of peroxisome proliferator-activated receptor γ and glucose transporter type 4. The results of the present study suggest that the phospholipid matrix used for the preparation of phytosomes critically influences their performance.

Anti-Inflammatory Effect of Ecklonia cava Extract on Porphyromonas gingivalis Lipopolysaccharide-Stimulated Macrophages and a Periodontitis Rat Model.

Ecklonia cava, an edible marine brown alga (Laminariaceae), is a rich source of phlorotannins. This study aimed to investigate the anti-inflammatory effect of Ecklonia cava ethanol extract (ECE, dieckol 10.6%, w/w) on Porphyromonas gingivalis lipopolysaccharide-stimulated inflammation in RAW 264.7 cells and in ligature-induced periodontitis in rats. The levels of nitric oxide (NO) and prostaglandin E2 were decreased by more than half on treatment with 100 µg/mL ECE. Downregulated tumor necrosis factor-α, interleukin (IL)-1ß, and IL-6 gene expression confirmed the anti-inflammatory properties of ECE. ECE treatment upregulated heme oxygenase-1 (HO-1) expression by 6.3-fold and increased HO-1/nuclear factor erythroid 2-related factor 2 (Nrf-2) signaling decreased nuclear factor-κB (NF-κB) translocation. ECE administration (400 mg/kg) significantly reduced gingival index, restricted tooth mobility, and prevented alveolar bone loss (p < 0.05). These beneficial effects were due to decreased inflammatory cell infiltration, IL-1ß production, and matrix metalloproteinase expression in gingival tissues. The ratio of receptor activator of nuclear factor-κB ligand (RANKL)/osteoprotegerin, a biomarker of periodontitis and osteolysis, was significantly decreased by ECE administration (p < 0.05). Thus, ECE has potential therapeutic effects for the alleviation of periodontal disease.

Hovenia dulcis Extract Attenuates High-Fat Diet-Induced Hepatic Lipid Accumulation and Hypertriglyceridemia in C57BL/6 Mice.

The effects of dietary supplementation with aqueous Hovenia dulcis Thunb. extract (HDE) (20 weeks) on high-fat diet (HFD)-induced nonalcoholic fatty liver disease and dyslipidemia were evaluated in mice. Supplementation with 200 and 800 mg/kg feed HDE (HDE200 and HDE800, respectively) resulted in no significant difference in growth in the HFD-fed groups. The triglyceride (TG) levels and free fatty acids were significantly decreased, whereas high-density lipoprotein cholesterol was increased in the HDE800 group (P < .05). The hepatic intracellular TGs were significantly decreased in the HDE-fed groups and lipogenic enzymes (acetyl CoA carboxylase, fatty acid synthase, stearoyl CoA desaturase, and diacylglycerol transferase) in the liver were significantly downregulated by HDE supplementation (P < .05). The diminished serum antioxidant enzyme activities in the HFD group were effectively restored by HDE supplementation, which also contributed to the attenuation of hyperlipidemia.

Ecklonia cava Extract Containing Dieckol Suppresses RANKL-Induced Osteoclastogenesis via MAP Kinase/NF-κB Pathway Inhibition and Heme Oxygenase-1 Induction.

Ecklonia cava, an edible marine brown alga (Laminariaceae), is a rich source of bioactive compounds such as fucoidan and phlorotannins. Ecklonia cava extract (ECE) was prepared using 70% ethanol extraction and ECE contained 67% and 10.6% of total phlorotannins and dieckol, respectively. ECE treatment significantly inhibited receptor activator of nuclear factor-κB ligand (RANKL)-induced osteoclast differentiation of RAW 264.7 cells and pit formation in bone resorption assay (p ＜0.05). Moreover, it suppressed RANKL-induced NF-κB and mitogen-activated protein kinase signaling in a dose dependent manner. Downregulated osteoclast-specific gene (tartrate-resistant acid phosphatase, cathepsin K, and matrix metalloproteinase-9) expression and osteoclast proliferative transcriptional factors (nuclear factor of activated T cells-1 and c-fos) confirmed ECE-mediated suppression of osteoclastogenesis. ECE treatment (100 µg/ml) increased heme oxygenase-1 expression by 2.5-fold and decreased intercellular reactive oxygen species production during osteoclastogenesis. The effective inhibition of RANKL-stimulated osteoclast differentiation and oxidative stress by ECE suggest that ECE has therapeutic potential in alleviating osteoclast-associated disorders.

Antiobesity Effect of Tricin, a Methylated Cereal Flavone, in High-Fat-Diet-Induced Obese Mice.

The antiobesity potential of tricin, a methylated cereal flavonoid, was examined using a high-fat-diet-induced obese mice model. The body weight ( P < 0.01) and body fat mass ( P < 0.05) were significantly decreased in the high-dose tricin supplementation group (TH: 200 mg/kg diet) in comparison to the high fat diet control group (CON) after a 12-week feeding trial. The serum (60.9 ± 2.09 mg/dL) and hepatic triglyceride levels (45.3 ± 4.42 nmol/mg protein) in the TH group were significantly decreased in comparison to the CON group (78.3 ± 5.09 mg/dL, 76.3 ± 8.10 nmol/mg protein), respectively. This antiobesity effect was attributed to a decrease in the expression of lipogenic markers crucial for fat synthesis in the liver (fatty acid synthase, stearoyl-CoA desaturase 1, elongation of long-chain fatty acids family member 6, glycerol-3-phosphate acyltransferase, and diglyceride acyltransferase) and suppressed expression of transcription factors associated with adipocyte differentiation (peroxisome proliferator-activated receptor γ and CCAAT/enhancer-binding protein α). These lipid-lowering effects are mediated by the activation of adenosine 5'-monophosphate-activated protein kinase.

Buffering Capacity of Dairy Powders and Their Effect on Yoghurt Quality.

Preheating conditions (low-, medium-, and high heat-) did not significantly affect the buffering capacity (BC) of skim milk powder (SMP), whereas the level of demineralization significantly affected the BC of whey powders (WP). Heat treatment (85°C for 30 min) of both SMP and WP (90% demineralized) mixtures (88:12, 76:24, 64:36 and 52:48; SMP:WP) resulted in a reduced BC, and the extent of this reduction increased with the proportion of WP increased in the samples. High-buffering milk prepared by the addition of phosphate salts (40 mM NaH2PO4 and 60 mM Na2HPO4) delayed the rate of pH decline during yoghurt fermentation. The high-buffering yoghurt showed a significantly higher water holding capacity (WHC) than that of control yoghurt (p<0.05), as well as a more uniform and interconnected microstructure with small pore sizes than those of control yoghurt. No significant differences were found between high-buffering and control yoghurt regarding the viable bacterial counts of starter. The manipulation BC can potentially improve the quality characteristics of yoghurts, such as WHC and texture.