Coagulation and proteolysis of high-protein milks in the gastric environment.

Gastric digestion of 2 commercial ultrafiltered milks and milk enriched with skim milk powder (to simulate concentration by reverse osmosis) was investigated and compared with the digestion of nonconcentrated milk. Curd formation and proteolysis of high-protein milks in simulated gastric conditions were studied using oscillatory rheology, extrusion testing, and gel electrophoresis. The presence of pepsin in the gastric fluid triggered coagulation at pH >6 and the elastic modulus of gels from high-protein milks was ~5 times larger than the gel from reference milk. Despite similar protein concentrations, the coagulum from milk enriched with skim milk powder showed higher resistance to shear deformation than the coagula from ultrafiltered milks. The gel structure was also more heterogeneous. During digestion, the degradation of coagula from high-protein milks was slowed down compared with the coagulum from reference milk, and intact milk proteins were still detected after 120 min. Differences in the digestion patterns of coagula from high-protein milks were observed and were associated with the proportion of minerals bound to caseins and the denaturation rate of whey proteins.

Tea catechin role in decreasing the oxidation of dairy beverages containing linseed oil.

Dairy beverages containing emulsified linseed oil is a suitable vehicle for delivering polyunsaturated fatty acids to consumers. However, these beverages are prone to oxidation. The purpose of this study was to evaluate the effect of adding various concentrations (0, 0.001, 0.01 and 0.1% (w/w)) of green tea extract (GTE) to dairy beverages (DB) containing linseed oil (2.0%, w/w), in order to inhibit lipid oxidation during storage at high temperature (50 °C) or under fluorescent light exposure. During storage, the concentration of catechin (C), epicatechin (EC) and epicatechin gallate (ECG) were significantly reduced (P ≤ 0.05) and degradation rate was greater when the DB were exposed to light (C 35%, EC 74% and ECG 68%) as compared to high temperature (C 34%, EC 45% and ECG 49%). In DB without GTE, the conjugated dienes (CD) hydroperoxides concentration increased significantly (P ≤ 0.05) from 23 mmol kg-1 fat to 243 mmol kg-1 fat under 6-day-light exposition, and to 83 mmol kg-1 fat under 6-day-heat temperature. The addition of GTE significantly increased the antioxidant capacity of DB and reduced the formation of CD, propanal and hexanal, induced by light exposure or high temperature. GTE at 0.10% completely inhibited CD formation during the storage period and reduced propanal and hexanal concentrations below the threshold.

Effect of emulsifiers on linseed oil emulsion structure, lipolysis and oxidation during in vitro digestion.

Health benefits have been associated with the consumption of omega-3 polyunsaturated fatty acids (PUFA). Linseed oil is rich in long chain omega-3 PUFA, but can generate toxic compounds due to its high susceptibility to oxidation. The nature of the emulsifier can affect both lipolysis and oxidation during digestion since these phenomena occur at the oil-water interface. The objective of this study was to compare the effect of low-molecular weight surfactants (cetyltrimethylammonium bromide (CTAB), Citrem), protein (sodium caseinate, fish gelatin) and polysaccharides (gum arabic, modified starch) on the structure of linseed oil emulsions, lipolysis and formation of reactive oxidation species during in vitro digestion. The emulsion stabilized with Citrem underwent extensive coalescence in the gastric phase, which strongly decreased the extent of lipid digestion and reduced the formation of oxidation markers relative to other emulsions. Emulsions stabilized by proteins and modified starch showed aggregation with partial coalescence in the gastric phase, but protein-stabilized emulsions showed better resistance to oxidation. This study shows that emulsifier properties affect the susceptibility of the emulsion to aggregation and coalescence in the gastrointestinal environment, and strongly influence the extent of lipid digestion and the formation of reactive oxidation products. These findings point out the importance of the choice of the emulsifier to control the lipid digestibility and the protection of sensible lipids thus promoting optimal nutritional properties in omega-3-enriched foods.

Antioxidant activity of milk and polyphenol-rich beverages during simulated gastrointestinal digestion of linseed oil emulsions.

Polyunsaturated fatty acids (PUFA) are associated with health benefits. However, high PUFA intake increases the risk of lipid oxidation and formation of potentially toxic lipid oxidation species. The objective of this study was to determine the antioxidant activity of milk fractions (whole milk, skim milk, acid whey, ultrafiltration (UF) permeate) and polyphenol-rich beverages (green tea, grape juice) during simulated gastrointestinal digestion. We also determined the effect of milk and polyphenol-rich beverages on the formation of advanced oxidation species during in vitro digestion of PUFA-rich emulsion. Antioxidant activity during digestion of milk fractions emphasized the important role of proteins (more specifically caseins) and the contribution of fat to the antioxidant capacity of milk. In comparison to milk, the antioxidant activity of polyphenol-rich beverages was at least four times higher. During digestion of a PUFA-rich emulsion, the formation of 4-hydroxyhexanal (4-HHE) and 4-hydroxynonenal (4-HNE) in the intestinal phase were respectively reduced by 60% and 75%, in the presence of milk or polyphenol-rich beverages. Further reduction was observed when the emulsion was co-digested with both, milk and polyphenol-rich beverages (89% for 4-HHE and 93% for 4-HNE). These results suggest that the combination of milk and polyphenol-rich beverages increases the antioxidant activity and synergistically reduces the formation of toxic lipid oxidation species during simulated digestion of PUFA-rich foods.

On the Use of Ultrafiltration or Microfiltration Polymeric Spiral-Wound Membranes for Cheesemilk Standardization: Impact on Process Efficiency.

Ultrafiltration (UF) and microfiltration (MF) are widely-used technologies to standardize the protein content of cheesemilk. Our previous work demonstrated that protein retention of a 0.1-µm MF spiral-wound membrane (SWM) was lower, but close to that of a 10 kDa UF one. Considering that the permeability of MF membranes is expected to be higher than that of UF ones, it was hypothesized that the former could improve the efficiency of the cheesemaking process. Consequently, the objectives of this work were to compare 0.1-µm MF and 10 kDa UF spiral-wound membranes in terms of (1) hydraulic and separation performance, (2) energy consumption and fouling behavior, (3) cheesemaking efficiency of retentates enriched with cream, and (4) economic performance in virtual cheesemaking plants. This study confirmed the benefits of using MF spiral-wound membranes to reduce the specific energy consumption of the filtration process (lower hydraulic resistance and higher membrane permeability) and to enhance the technological performance of the cheesemaking process (higher vat yield, and protein and fat recoveries). However, considering the higher serum protein retention of the UF membrane and the low price of electricity in Canada, the UF scenario remained more profitable. It only becomes more efficient to substitute the 10 kDa UF SWM by the 0.1-µm MF when energy costs are substantially higher.

Identification of texture parameters influencing commercial cheese matrix disintegration and lipid digestion using an in vitro static digestion model.

Cheese characteristics, such as composition or textural properties, can impact the matrix degradation rate which could modulate the bioaccessibility of fatty acids during digestion. The aim of this study was to identify texture parameters influencing cheese degradation in a gastrointestinal environment. A static in vitro digestion model has been used on nine commercial cheeses: young and aged cheddar, regular and light cream cheese, parmesan, feta, camembert, mozzarella, and sliced processed cheese. At the end of gastric digestion, camembert and mozzarella presented the lowest matrix disintegration whereas aged cheddar, regular and light cream cheeses showed the highest. For all cheeses, the fatty acid release was fast during the first 30 min of duodenal digestion and slowed down afterwards. A partial least square regression revealed that springiness, cohesiveness, and hardness were negatively correlated to the rate of cheese disintegration during gastric digestion. In addition, textural parameters were not correlated with free fatty acid release. By modulating cheese texture, it could be possible to influence matrix disintegration during gastrointestinal digestion which could have an impact on lipids release.

Effect of milk proteins and food-grade surfactants on oxidation of linseed oil-in-water emulsions during in vitro digestion.

Health benefits are associated with polyunsaturated fatty acids, but their sensitivity to oxidation may generate toxic oxidation species. The objective of this study was to compare the effect of milk proteins (casein, whey protein) and surfactants (Citrem, Tween 20) on the in vitro digestion and oxidation of linseed oil emulsions. The emulsion produced with Tween 20 resisted coalescence in the gastric phase and showed the highest concentrations of free fatty acids and reactive carbonyl compounds in the intestinal digestion phase. The Citrem-stabilized emulsion showed extensive coalescence in the gastric environment, which reduced lipolysis and the formation of advanced oxidation species. The protein-stabilized emulsions showed aggregation with some coalescence in the gastric phase, and casein provided better protection than whey protein against oxidation. This study suggests that the mechanism of emulsion destabilization in the gastric environment and the type of protein can modulate lipolysis and oxidation during in vitro digestion.

Reprint of "Postprandial lipemia and fecal fat excretion in rats is affected by the calcium content and type of milk fat present in Cheddar-type cheeses".

The aim of this study was to better understand the effect of calcium on the bioavailability of milk lipids from a cheese matrix using a rat model. Cheddar-type cheeses were manufactured with one of three types of anhydrous milk fat, control, olein or stearin, and salted with or without CaCl2. The cheeses were fed to rats and postprandial lipemia was monitored. Feces were analyzed to quantify fatty acids excreted as calcium soaps. Higher calcium concentration in cheese caused a higher and faster triacylglycerol peak in blood, except for cheeses containing stearin. Furthermore, calcium soaps were more abundant in feces when the ingested cheese had been enriched with calcium and when the cheese was prepared with stearin. Increased lipid excretion was attributable to the affinity of saturated long-chain fatty acids for calcium. Results showed that lipid bioaccessibility can be regulated by calcium present in Cheddar cheese. This study highlights the nutritional interaction between calcium and lipids present in the dairy matrix and confirms its physiological repercussions on fatty acid bioavailability.

Evaluation of casein as a binding ligand protein for purification of alpha-lactalbumin from beta-lactoglobulin under high hydrostatic pressure.

Fractionation of ß-lactoglubulin (ß-lg) and α-lactalbumin (α-la) using conventional separation technologies remains challenging mainly due to similar molecular weight. Herein, casein (CN) was used as ligand protein to specifically aggregate ß-lg under high hydrostatic pressure (HHP) in order to separate α-la after acidification to pH 4.6. Specifically, we studied the effect of different concentration of CN on α-la purity and recovery. Model solutions of α-la, ß-lg and CN (from 0 to 5 mg/mL) were pressurized (600 MPa-5 min). After acidification and centrifugation of pressure-treated solutions, purity of α-la was increased up to 78% with a recovery of 88% for solution without CN. In contrast with our initial hypothesis, the presence of CN decreased ß-lg pressure-induced aggregation and co-precipitation upon acidification and significantly reduced purity (∼71%). Therefore, our results suggest a chaperone-like activity of CN on ß-lg pressure-induced aggregation which needs further investigation.

Postprandial lipemia and fecal fat excretion in rats is affected by the calcium content and type of milk fat present in Cheddar-type cheeses.

The aim of this study was to better understand the effect of calcium on the bioavailability of milk lipids from a cheese matrix using a rat model. Cheddar-type cheeses were manufactured with one of three types of anhydrous milk fat, control, olein or stearin, and salted with or without CaCl2. The cheeses were fed to rats and postprandial lipemia was monitored. Feces were analyzed to quantify fatty acids excreted as calcium soaps. Higher calcium concentration in cheese caused a higher and faster triacylglycerol peak in blood, except for cheeses containing stearin. Furthermore, calcium soaps were more abundant in feces when the ingested cheese had been enriched with calcium and when the cheese was prepared with stearin. Increased lipid excretion was attributable to the affinity of saturated long-chain fatty acids for calcium. Results showed that lipid bioaccessibility can be regulated by calcium present in Cheddar cheese. This study highlights the nutritional interaction between calcium and lipids present in the dairy matrix and confirms its physiological repercussions on fatty acid bioavailability.

Differential impact of the cheese matrix on the postprandial lipid response: a randomized, crossover, controlled trial.

Background: In a simulated gastrointestinal environment, the cheese matrix modulates dairy fat digestion. However, to our knowledge, the impact of the cheese matrix on postprandial lipemia in humans has not yet been evaluated.Objective: In healthy subjects, we compared the impact of dairy fat provided from firm cheese, soft cream cheese, and butter on the postprandial response at 4 h and on the incremental area under the curve (iAUC) of plasma triglycerides.Design: Forty-three healthy subjects were recruited to this randomized, crossover, controlled trial. In random order at intervals of 14 d and after a 12-h fast, subjects ingested 33 g fat from a firm cheese (young cheddar), a soft cream cheese (cream cheese), or butter (control) incorporated into standardized meals that were matched for macronutrient content. Plasma concentrations of triglycerides were measured immediately before the meal and 2, 4, 6, and 8 h after the meal.Results: Cheddar cheese, cream cheese, and butter induced similar increases in triglyceride concentrations at 4 h (change from baseline: +59%, +59%, and +62%, respectively; P = 0.9). No difference in the triglyceride iAUC0-8 h (P-meal = 0.9) was observed between the 3 meals. However, at 2 h, the triglyceride response caused by the cream cheese (change from baseline: +44%) was significantly greater than that induced by butter (change from baseline: +24%; P = 0.002) and cheddar cheese (change from baseline: +16%; P = 0.0004). At 6 h, the triglyceride response induced by cream cheese was significantly attenuated compared with that induced by cheddar cheese (change from baseline: +14% compared with +42%; P = 0.0004).Conclusion: This study demonstrates that the cheese matrix modulates the impact of dairy fat on postprandial lipemia in healthy subjects. This trial was registered at clinicaltrials.gov as NCT02623790.

Physical, microbiological and rheological properties of probiotic yogurt supplemented with grape extract.

In this study, yogurt was supplemented with 1.5 and 3.0 g L-1 of grape extract, inoculated culture containing Streptococcus thermophilus, Lactobacillus bulgaricus, Lactobacillus acidophilus and Bifidobacterium bb12 bifidum, fermented and stored at 4 °C. Acid production, microbial growth, gel strength, syneresis, rheological and sensory properties were studied. An increase in grape extract concentration extended fermentation time. Bacterial strains were found in at least 109 CFU100 g-1 of yogurt showing the possibility of probiotic yogurt production with grape extract. Gel strength decreased with increasing concentration of grape extract while syneresis increased. The addition of grape extract changed the dilatant behavior to a pseudoplastic behavior, decreased yield stress, whereas k values increased. Sensory attributes (color, flavor, taste, texture and appearance) didn't differ significantly.

Effect of denatured whey protein concentrate and its fractions on cheese composition and rheological properties.

The objectives of this study were (1) to assess the effect of a denatured whey protein concentrate (DWPC) and its fractions on cheese yield, composition, and rheological properties, and (2) to separate the direct effect of the DWPC or its fractions on cheese rheological properties from the effect of a concomitant increase in cheese moisture. Semihard cheeses were produced at a laboratory scale, and mechanical properties were characterized by dynamic rheometry. Centrifugation was used to induce a moisture gradient in cheese to separate the direct contribution of the DWPC from the contribution of moisture to cheese mechanical properties. Cheese yield increased and complex modulus (G*) decreased when the DWPC was substituted for milk proteins in milk. For cheeses with the same moisture content, the substitution of denatured whey proteins for milk proteins had no direct effect on rheological parameters. The DWPC was fractionated to evaluate the contribution of its different components (sedimentable aggregates, soluble component, and diffusible component) to cheese yield, composition, and rheological properties. The sedimentable aggregates were primarily responsible for the increase in cheese yield when DWPC was added. Overall, moisture content explained to a large extent the variation in cheese rheological properties depending on the DWPC fraction. However, when the effect of moisture was removed, the addition of the DWPC sedimentable fraction to milk increased cheese complex modulus. Whey protein aggregates were hypothesized to act as active fillers that physically interact with the casein matrix and confer rigidity after pressing.

Influence of dairy matrices on nutrient release in a simulated gastrointestinal environment.

The objective of this study was to compare the kinetics of the release of nutrients (peptides and fatty acids) from different dairy matrices (milks, yogurts, and cheeses) in a simulated gastrointestinal environment. Prior to processing, different heat and homogenization treatments were applied to milks, and different drainage pH levels were used to control calcium concentration in cheeses. The dairy matrices were then subjected to simulated digestion. Matrix degradation, protein hydrolysis, and fat hydrolysis were analyzed during the gastric and intestinal digestion phases. Intense heat treatment of milk induced faster digestion of proteins in the gastric environment. Cheeses were more resistant to protein and lipid digestion than liquid or semi-solid matrices were. No direct relationship could be established between disintegration kinetics and cheese rheological properties. Fatty acid release in the intestinal phase was much faster when matrices were produced from homogenized milk. For cheeses, greater fatty acid release could not be related to faster matrix disintegration, suggesting that the lipid droplet size dispersion was more important than matrix breakdown was for the modulation of lipid digestion kinetics. Calcium soaps were produced in the intestinal environment, and their concentration was higher during the digestion of cheeses in comparison with milks and yogurts. These results suggest that processing-induced modifications to the composition, microstructure, and rheological properties of dairy matrices could be used to control nutrient delivery.

Effect of calcium on fatty acid bioaccessibility during in vitro digestion of Cheddar-type cheeses prepared with different milk fat fractions.

Calcium plays an important role in intestinal lipid digestion by increasing the lipolysis rate, but also limits fatty acid bioaccessibility by producing insoluble Ca soaps with long-chain fatty acids at intestinal pH conditions. The aim of this study was to better understand the effect of Ca on the bioaccessibility of milk fat from Cheddar-type cheeses. Three anhydrous milk fats (AMF) with different fatty acid profiles (olein, stearin, or control AMF) were used to prepare Cheddar-type cheeses, which were then enriched or not with Ca using CaCl2 during the salting step. The cheeses were digested in vitro, and their disintegration and lipolysis rates were monitored during the process. At the end of digestion, lipids were extracted under neutral and acidic pH conditions to compare free fatty acids under intestinal conditions in relation to total fatty acids released during the digestion process. The cheeses prepared with the stearin (the AMF with the highest ratio of long-chain fatty acids) were more resistant to disintegration than the other cheeses, owing to the high melting temperature of that AMF. The Ca-enriched cheeses had faster lipolysis rates than the regular Ca cheeses. Chromatographic analysis of the digestion products showed that Ca interacted with long-chain fatty acids, producing Ca soaps, whereas no interaction with shorter fatty acids was detected. Although higher Ca levels resulted in faster lipolysis rates, driven by the depletion of reaction products as Ca soaps, such insoluble compounds are expected to reduce the bioavailability of fatty acids by hindering their absorption. These effects on lipid digestion and absorption are of interest for the design of food matrices for the controlled release of fat-soluble nutrients or bioactive molecules.

Antimicrobial activity of buttermilk and lactoferrin peptide extracts on poultry pathogens.

Antibiotics are commonly used in poultry feed as growth promoters. This practice is questioned given the arising importance of antibiotic resistance. Antimicrobial peptides can be used as food additives for a potent alternative to synthetic or semi-synthetic antibiotics. The objective of this study was to develop a peptide production method based on membrane adsorption chromatography in order to produce extracts with antimicrobial activity against avian pathogens (Salmonella enterica var. Enteritidis, Salmonella enterica var. Typhimurium, and two Escherichia coli strains, O78:H80 and TK3 O1:K1) as well as Staphylococcus aureus. To achieve this, buttermilk powder and purified lactoferrin were digested with pepsin. The peptide extracts (<10 kDa) were fractionated depending on their charges through high-capacity cation-exchange and anion-exchange adsorptive membranes. The yields of cationic peptide extracts were 6·3 and 15·4% from buttermilk and lactoferrin total peptide extracts, respectively. Antimicrobial activity was assessed using the microdilution technique on microplates. Our results indicate that the buttermilk cationic peptide extracts were bactericidal at less than 5 mg/ml against the selected avian strains, with losses of 1·7 log CFU/ml (Salm. Typhimurium) to 3 log CFU/ml (E. coli O78:H80); viability decreased by 1·5 log CFU/ml for Staph. aureus, a Gram-positive bacterium. Anionic and non-adsorbed peptide extracts were inactive at 5 mg/ml. These results demonstrate that membrane adsorption chromatography is an effective way to prepare a cationic peptide extract from buttermilk that is active against avian pathogens.

Effect of the consumption of ß-lactoglobulin and epigallocatechin-3-gallate with or without calcium on glucose tolerance in C57BL/6 mice.

Interactions between ß-lactoglobulin (ß-lg) and epigallocatechin-3-gallate (EGCG) may modulate their health benefits. The objective of this study was therefore to investigate the synergistic effect of consuming ß-lg and EGCG complexes on glucose tolerance of C57BL/6 male mice given an oral glucose tolerance test (OGTT) and randomized to one of the following treatments administered prior to the OGTT: 1) simulated milk ultrafiltrate (SMUF(-)), 2) SMUF(-) + EGCG, 3) SMUF(-) + ß-lg, 4) SMUF(-) + EGCG + ß-lg, 5) SMUF + calcium (SMUF(+)) and 6) SMUF(+) + EGCG + ß-lg. We found no significant between-group difference in postprandial glucose response. However, when mice were separated in those who received ß-lg from those who did not, we found that the latter displayed significantly higher postprandial glucose concentrations. Our results support the beneficial impact of ß-lg on glycemic control and suggest that concomitant EGCG or calcium consumption does not improve this effect.

Antioxidant activity and nutrient release from polyphenol-enriched cheese in a simulated gastrointestinal environment.

Green tea polyphenols are recognized for their antioxidant properties and their effects on lipid digestion kinetics. Polyphenols are sensitive to degradation in the intestinal environment. Interactions with dairy proteins could modulate the stability and biological activity of polyphenols during digestion. The objective of this study was to evaluate the release of nutrients (polyphenols, fatty acids and peptides) and the antioxidant activity in polyphenol-enriched cheese containing different levels of calcium in a simulated gastrointestinal environment. The relationship between cheese matrix texture, matrix degradation and nutrient release during digestion was also studied. Green tea extract was added to milk at 0% or 0.1%, and cheeses were produced on a laboratory scale. The level of available calcium was adjusted to low (Ca(low)), regular (Ca(reg)) or high (Ca(high)) during the salting step of the cheese-making process. Cheeses were subjected to simulated digestion. The rate and extent of fatty acid release were 21% lower for Ca(low) cheese than for Ca(reg) and Ca(high) cheeses. The greater adhesiveness of Ca(low) cheese, which resulted in lower rates of matrix degradation and proteolysis, contributed to the reduced rate of lipolysis. The presence of green tea extract in cheese reduced the release of free fatty acids at the end of digestion by 7%. The addition of green tea extract increased cheese hardness but did not influence matrix degradation or proteolysis profiles. The formation of complexes between tea polyphenols and proteins within the cheese matrix resulted in a more than twofold increase in polyphenol recovery in the intestinal phase compared with the control (tea polyphenol extract incubated with polyphenol-free cheese). Antioxidant activity was 14% higher in the digest from polyphenol-enriched cheese than in the control. These results suggest that cheese is an effective matrix for the controlled release of nutrients and for the protection of green tea polyphenol integrity and biological activity in the gastrointestinal environment.

Protein-Protein Multilayer Oil-in-Water Emulsions for the Microencapsulation of Flaxseed Oil: Effect of Whey and Fish Gelatin Concentration.

The impact of whey protein isolate (WPI) and fish gelatin (FG) deposited sequentially at concentrations of 0.1, 0.5, and 0.75% on the surface of primary oil-in-water emulsions containing 5% flaxseed oil stabilized with either 0.5% fish gelatin or whey protein, respectively, was investigated. The results revealed that the adsorption of WPI/FG or FG/WPI complexes to the emulsion interface led to the formation of oil-in-water (o/w) emulsions with different stabilities and different protection degrees of the flaxseed oil. Deposition of FG on the WPI primary emulsion increased the particle size (from 0.53 to 1.58 µm) and viscosity and decreased electronegativity (from -23.91 to -11.15 mV) of the complexes. Different trends were noted with the deposition of WPI on the FG primary emulsion, resulting in decreasing particle size and increasing electronegativity and viscosity to a lower extent. Due to the superior tension-active property of WPI, the amount of protein load in the WPI primary emulsion as well as in WPI/FG complex was significantly higher than the FG counterparts. A multilayer emulsion made with 0.5% WPI/0.75% FG exhibited the lowest oxidation among all of the multilayered emulsions tested (0.32 ppm of hexanal) after 21 days, likely due to the charge effect of FG that may prevent pro-oxidant metals to interact with the flaxseed oil.