Microstructural studies of imitation cheese with a shift in continuous phase.

When casein is replaced with starch in imitation cheese, the functionality changes. Three different microscopy methods were applied to understand the microstructural differences in the product depending on which component dominates the microstructure. Confocal Laser Scanning Microscopy (CLSM) for component identification. Scanning Electron Microscopy (SEM) and Cryogenic Scanning Electron Microscopy (Cryo-SEM) for studying surface structures. Differences in the surface structures were detected between SEM and Cryo-SEM. In SEM, starch appeared rough and protein smooth, while in Cryo-SEM no starch roughness of the surface was found. A change in starch modification and effects of protein prehydration was also analysed. Adding octenyl succinic anhydride (OSA) modified starch for emulsifying properties resulted in a microstructure with fragmented protein at a protein level of 7 %, but not at 9 or 12 %. Protein prehydration had limited effect on microstructure. On a macrostructural level, the change to an emulsifying starch increased hardness in imitation cheese with 7 and 9 % protein. Protein prehydration slightly decreased the hardness, but the difference was not significant at all concentrations. This research provides valuable information about the microstructure of imitation cheese at a 50/50 composition, how the microstructure changes with an emulsifying starch and what occurs after a protein prehydration was included in the production.

Lupin protein-stabilized oil droplets contribute to structuring whey protein emulsion-filled gels.

This work aimed to understand the role of lupin protein or mixed lupin-whey protein stabilized oil droplets on the texture and microstructure of a heat-induced whey protein gel. Protein-stabilized emulsions were compared to surfactant-stabilized emulsions to investigate the potential of their interfacial interactions to impart unique structures in the filled gels. The structure development was followed in situ using rheology and the final heat-induced gels were characterized by small and large amplitude oscillatory rheology and confocal microscopy. The development of the gel modulus as well as the final gel properties were linked to the type of interactions between the whey protein matrix and the protein adsorbed at the oil interface. The final gels were selectively dissolved in various buffers, and the results showed that replacing interfacial whey protein with lupin protein resulted in a reduced amount of disulfide bridges, explaining the softer gel in the lupin containing gels compared to those with whey protein. Non-covalent interactions were the main forces involved in the formation of actively filled droplets in the gel network. This work demonstrated that by modulating the interfacial composition of the oil droplets, differing gel structures could be achieved due to differences in the protein-protein interactions between the continuous and the interfacial phase. There is therefore potential for the development of innovative products using lupin-whey protein mixtures, by careful control of the processing steps and the matrix composition.

Role of the pea protein aggregation state on their interfacial properties.

HYPOTHESIS: Plant protein ingredients from similar sources can vary in functionality not only because of compositional differences, but also because of differences in their structure depending on their processing history. It is essential to understand these distinctions to develop novel food emulsion using plant proteins. It is hypothesized that differing interfacial properties can be attributed to their structures, aggregation, and colloidal states. EXPERIMENTS: The adsorption behavior of a commercial protein isolate, homogenized or non-homogenized, was compared to a mildly extracted isolate to evaluate the effect of aggregation state and structural differences. After characterization of the particle size and protein composition, the interfacial properties were compared. FINDINGS: Atomic force microscopy provided evidence of interfaces packed with protein oligomers regardless of the treatment. Differences in adsorption kinetics and interfacial shear rheology depending on oil polarity suggested different interfacial structures. A polydisperse mixture of protein oligomers resulted in increased rearrangements and protein-protein interactions at the interface. Homogenization of commercial proteins resulted in a lower interfacial tension and less elastic interfaces compared to those of native proteins due to the presence of larger aggregates. This study highlights how the interfacial properties can be related to the protein aggregation state resulting from differences in processing history.

Correction: Hammershøj et al. Dual-Purpose Poultry in Organic Egg Production and Effects on Egg Quality Parameters. Foods 2021, 10, 897.

The authors would like to make the following correction to the published paper [...].

Effect of Dairy Matrix on the Postprandial Blood Metabolome.

This study investigated the postprandial plasma metabolome following consumption of four dairy matrices different in texture and structure: cheddar cheese (Cheese), homogenized cheddar cheese (Hom. Cheese), and micellar casein isolate (MCI) with cream (MCI Drink) or a MCI Gel. An acute, randomized, crossover trial in male participants (n = 25) with four test days was conducted. Blood samples were collected during an 8-h postprandial period after consumption of a meal similar in micro- and macronutrients containing one of the four dairy matrices, and the metabolome was analyzed using nuclear magnetic resonance (NMR) spectroscopy. A liquid dairy matrix (MCI Drink) resulted in a faster absorption of amino acids compared to products, representing either a semi-solid (MCI Gel and Hom. Cheese) or solid (Cheese) dairy matrix. For the MCI Gel, plasma concentration of acetic acid and formic acid increased approximately 2 h following consumption, while 3-hydroxybyturate and acetoacetic acid increased approximately 6 h after consumption. The structure and texture of the dairy matrix affected the postprandial absorption of amino acids, as revealed by the plasma metabolome. Our study furthermore pointed at endogenous effects associated with consumption of dairy products containing glucono-δ-lactone.

Biorefinery of Green Biomass─How to Extract and Evaluate High Quality Leaf Protein for Food?

There is a growing need for protein for both feed and food in order to meet future demands. It is imperative to explore and utilize novel protein sources such as protein from leafy plant material, which contains high amounts of the enzyme ribulose-1,5-biphosphate carboxylase/oxygenase (RuBisCo). Leafy crops such as grasses and legumes can in humid climate produce high protein yields in a sustainable way when compared with many traditional seed protein crops. Despite this, very little RuBisCo is utilized for foods because proteins in the leaf material has a low accessibility to monogastrics. In order to utilize the leaf protein for food purposes, the protein needs to be extracted from the fiber rich leaf matrix. This conversion of green biomass to valuable products has been labeled green biorefinery. The green biorefinery may be tailored to produce different products, but in this Review, the focus is on production of food-grade protein. The existing knowledge on the extraction, purification, and concentration of protein from green biomass is reviewed. Additionally, the quality and potential application of the leaf protein in food products and side streams from the green biorefinery will be discussed along with possible uses of side streams from the protein production.

Matrix structure of dairy products results in different postprandial lipid responses: a randomized crossover trial.

BACKGROUND: The dairy matrix may influence digestion and absorption of lipids and thereby risk of cardiovascular diseases (CVDs). However, few postprandial studies have compared dairy products that differed only in terms of their matrix. OBJECTIVES: We aimed to investigate acute 8-h postprandial lipid, glycemic, and appetite responses after intake of isoenergetic dairy meals with different matrixes, but similar nutritional composition. METHODS: Twenty-five normal-weight men (18-40 y old) were enrolled in a randomized controlled crossover trial. On 4 test days, a meal with 1 of 4 dairy products was served: cheddar cheese (Cheese), homogenized Cheese (Hom. Cheese), micellar casein isolate (MCI) with cream (MCI Drink), and a gel produced from the MCI Drink by addition of Glucono Delta-Lactone (MCI Gel). The fat- and protein-matched dairy products differed in terms of their casein network, fat droplet size, and/or texture. Blood biochemistry and appetite responses were collected. RESULTS: Eighteen participants completed the trial. Postprandial triglycerides (TGs) (primary outcome) increased by (mean ± SEM) 0.24 ± 0.07 and 0.19 ± 0.07 mmol/L after MCI Gel compared with Cheese and Hom. Cheese, respectively (both P ≤ 0.05). Likewise, MCI Gel increased TG incremental AUC compared with Cheese and Hom. Cheese (both P < 0.05), and peak compared with Cheese (P < 0.05). ApoB-48 (primary outcome) was unaffected by dairy matrix. For free fatty acids (FFAs), glucose, and insulin, time × meal interactions were observed (all P < 0.001). During the first 2 h, FFAs were lower for Cheese than for MCI products, whereas the opposite was observed for glucose and insulin. CONCLUSIONS: Postprandial TG but not apoB-48 response was higher after MCI Gel, indicating that the type of casein network influences lipid responses. This suggests that the dairy matrix may also affect risk factors for CVDs. Reducing fat droplet size (i.e., Hom. Cheese) did not affect blood biochemistry.This trial was registered at clinicaltrials.gov as NCT03656367.

Progression of Postprandial Blood Plasma Phospholipids Following Acute Intake of Different Dairy Matrices: A Randomized Crossover Trial.

Studies have indicated that the dairy matrix can affect postprandial responses of dairy products, but little is known about the effect on postprandial plasma phospholipid levels. This study investigated postprandial plasma phospholipid levels following consumption of four different dairy products that are similar in micro and macro nutrients, but different in texture and structure: cheddar cheese (Cheese), homogenized cheddar cheese (Hom. Cheese), micellar casein isolate with cream (MCI Drink) or a gel made from the MCI Drink (MCI Gel). The study was an acute randomized, crossover trial in human volunteers with four test days. Blood samples were collected during an 8 h postprandial period and the content of 53 plasma phospholipids was analysed using liquid chromatography-mass spectrometry (LC-MS). No meal-time interactions were revealed; however, for nine of the 53 phospholipids, a meal effect was found. Thus, the results indicated a lower plasma level of specific lyso-phosphatidylethanolamines (LPEs) and lyso-phosphatidylcholines (LPCs) following consumption of the MCI Gel compared to the MCI Drink and Hom. Cheese, which might be attributed to an effect of viscosity. However, further studies are needed in order to reveal more details on the effect of the dairy matrix on postprandial phospholipids.

Dual-Purpose Poultry in Organic Egg Production and Effects on Egg Quality Parameters.

Egg laying genotypes have been selected for generations due to their high yield and egg quality, resulting in efficient feed utilization and low body weight; hence, they are not suitable for meat production. This imposes an issue for the male layer chicks, which are killed at one day old. Because of ethical and food waste concerns, the search for suitable dual-purpose genotypes in order to avoid euthanasia of male day-old chicks has intensified. The aim of the present study is to evaluate potential dual-purpose genotypes for their egg quality compared to a representative egg laying genotype. Two dual-purpose genotypes with divergent characteristics were evaluated: genotype A represented an experimental crossbreed based on a broiler type male and an egg layer female, and genotype C was a crossbreed of a layer type. These were compared to a rustic genotype B and a control genotype D, which was an egg layer. Eggs were collected six times during the period of 21­54 weeks of hen age, i.e., a total of 990 shell eggs were analyzed. Examined parameters were weights of egg, shell, yolk, and albumen, by calculating their relative proportions. Shell quality was assessed by shell strength, shell stiffness, and shell thickness. Yolk quality was determined as yolk color and inclusions of blood and meat spots, and albumen quality was evaluated in terms of pH and dry matter (DM) content. The egg layer genotype produced the smallest eggs with least blood and meat spot inclusions compared to that produced by the three dual-purpose genotypes. Shell quality was superior for the layer genotype. However, the experimental genotype A laid eggs of comparable shell quality, albumen DM, and yolk weight, but also with the darkest and most red-yellow colored yolk. The two other dual-purpose genotypes produced eggs of low-medium quality. In conclusion, the genotype A could serve as dual-purpose genotype from an egg quality perspective.

Mechanism behind the degradation of aqueous norbixin upon storage in light and dark environment.

Buffered aqueous solutions of norbixin were stored in light and dark, and analyzed using mass spectrometry. Compounds with both higher and lower masses than norbixin were detected, suggesting the formation of oxidation products and oxidative cleavage products of norbixin. The norbixin oxidation products included compounds containing several oxidations. The amounts of oxidation products of norbixin increased during storage in both light and dark, but in light, the development accelerated. Scavengers of superoxide radical anion (superoxide dismutase), hydrogen peroxide (catalase), hydroxyl radicals (mannitol) and singlet oxygen (sodium azide) and carbon-centered radicals (DMPO) were tested to determine if any of the reactive species were involved in the degradation of norbixin. Of these, only DMPO decreased the bleaching of norbixin indicating the involvement of carbon-centered radicals. Multiple oxidations of norbixin might be a result of a radical chain reaction involving peroxyl and carbon-centered radicals even though not detectable with electron spin resonance.

A Comprehensive Approach to Assess Feathermeal as an Alternative Protein Source in Aquafeed.

The effect of partially replacing fishmeal in aquafeed with feathermeal (FTH) at three levels (0%: FTH0, 8%: FTH8, 24%: FTH24) and two extrusion temperatures (100 and 130 °C) was evaluated in rainbow trout (Oncorhynchus mykiss) with respect to growth performance, metabolism response, and oxidative status of the feed proteins. Multivariate data analyses revealed that FTH24 correlated positively with high levels of oxidation products, amino acids (AA) racemization, glucogenic AAs level in liver, feed intake (FI), specific growth rate (SGR), and feed conversion ratio (FCR); and low AAs digestibility. Both FI and SGR were significantly increased when 8 and 24% feathermeal was included in the feed extruded at 100 °C, while there was a negative effect on FCR in fish fed FTH24. In conclusion, higher oxidation levels in FTH24 may give rise to metabolic alterations while lower levels of FTH may be considered as fishmeal substitute in aquafeed for rainbow trout.

Appearance and Textural Properties of Sheared Low Concentration Potato Protein Gels-Impact of Drying Method, pH, and Ionic Strength.

The objective was to prepare sheared gels of potato protein concentrate and evaluate the effect of pH (3, ∼4, ∼7), ionic strength (15 or 200 mM) and protein drying conditions (spray or freeze drying) on the final appearance and rheological characteristics. Heat-set gels 3 % (w/w) at a high ionic strength (200 mM) resulted in an inhomogeneous appearance with presence of clots, while low ionic strength (15 mM) gave homogenous structures. Gels prepared at pH 3 became transparent while preparation above pH 3.0 resulted in high turbidity. Heat treatment and cooling resulted in gelation for all samples except freeze dried powder at pH 3.0. Flow curves during shear from 0.1 to 100 s-1 were fitted by the Herschel-Bulkley model indicating shear thinning behavior for all samples except the freeze dried sample at pH 3 which displayed a Newtonian behavior. Oscillatory measurements after shear indicated viscus behavior (phase angle above 45°) for the spray dried sample at pH 3, and gelled behavior (phase angle above 45°) for the remaining gelled samples. Structure recovery was observed after shear in all samples except at pH 3.0. The data shows potato protein can be used as ingredient in protein beverages.

Impact of triacylglycerol composition on shear-induced textural changes in highly saturated fats.

This study demonstrates a strong interaction between triacylglycerol (TAG) composition and effects of shear rate on the microstructure and texture of fats. Cocoa butter alternatives with similar saturated fat content, but different major TAGs (PPO-, PSO-, SSO-, POP- and SOS-rich blends) were evaluated. Results show how shear can create a harder texture in fat blends based on symmetric monounsaturated TAGs (up to ∼200%), primarily due to reduction in crystal size, whereas shear has little effect on hardness of asymmetric monounsaturated TAGs. Such differences could not be ascribed to differences in the degree of supercooling, but was found to be a consequence of differences in the crystallisation behaviour of different TAGs. The fractal dimension was evaluated by dimensional detrended fluctuation analysis and Fourier transformation of microscopy images. However, the concept of fractal patterns was found to be insufficient to describe microstructural changes of fat blends with high solid fat content.

Protein denaturation of whey protein isolates (WPIs) induced by high intensity ultrasound during heat gelation.

In this study, the impact of high intensity ultrasound (HIU) on proteins in whey protein isolates was examined. Effects on thermal behavior, secondary structure and nature of intra- and intermolecular bonds during heat-induced gelling were investigated. Ultrasonication (24 kHz, 300 W/cm(2), 2078 J/mL) significantly reduced denaturation enthalpies, whereas no change in secondary structure was detected by circular dichroism. The thiol-blocking agent N-ethylmaleimide was applied in order to inhibit formation of disulfide bonds during gel formation. Results showed that increased contents of α-lactalbumin (α-La) were associated with increased sensitivity to ultrasonication. The α-La:ß-lactoglobulin (ß-Lg) ratio greatly affected the nature of the interactions formed during gelation, where higher amounts of α-La lead to a gel more dependent on disulfide bonds. These results contribute to clarifying the mechanisms mediating the effects of HIU on whey proteins on the molecular level, thus moving further toward implementing HIU in the processing chain in the food industry.

Impact of NaCl reduction in Danish semi-hard Samsoe cheeses on proliferation and autolysis of DL-starter cultures.

Reduction of sodium chloride (NaCl) in cheese manufacturing is a challenge for the dairy industry. NaCl has a profound role on microbial development influencing cheese sensory and technological properties. The purpose of this work was to investigate how proliferation, distribution and autolysis of two commercial DL-starter cultures (C1 and C2) used in the production of Danish semi-hard Samsoe cheeses were affected by reduced NaCl levels. Cheeses containing <0.3% (unsalted), 2.3% (reduced-salt) and 3.4% (normal-salted) (w/v) NaCl in moisture were produced and analyzed during 12 weeks of ripening. Lactic acid bacteria (LAB), distribution of bacteria as single cells or microcolonies, their viability in the cheeses and cell autolysis were monitored during ripening, as well as the impact of NaCl content and autolysis on the formation of free amino acids (FAA). Reduction of NaCl resulted in higher LAB counts at the early stages of ripening, with differences between the two DL-starter cultures. The unsalted cheeses produced with C1 had retained a significantly higher number of the initial LAB counts (cfu/g) after 1 and 2 weeks of ripening (i.e. 58% and 71%), compared to the normal-salted cheeses (i.e. 22% and 21%), whereas no significant difference was found between the reduced-salt (i.e. 31% and 35%) and normal-salted cheeses. At the later stages of ripening (i.e. 7 and 11 weeks) NaCl had no significant influence. For cheeses produced with C2, a significant influence of NaCl was only found in cheeses ripened for 7 weeks, where the unsalted and reduced-salt cheeses had retained a significantly higher number of the initial LAB counts (cfu/g) (i.e. 39% and 38%), compared to the normal-salted cheeses (i.e. 21%). In the Samsoe cheeses, bacteria were organized as single cells, in groups of 2-3 cells or in groups of ≥4 cells. During ripening the decrease in the number of viable bacteria was mainly due to a reduction in the number of viable bacteria organized in groups of ≥4 cells. A negative correlation between NaCl content and PepX activity was observed. At the end of ripening the total FAA content was lower in the unsalted cheeses, compared to the reduced- and normal-salted cheeses. In conclusion, NaCl had a significant influence on proliferation of both DL-starter cultures. However, the influence of NaCl on culture development was more pronounced in cheeses produced with DL-starter culture C1. As both texture and taste are parameters known to be affected by the development of the starter culture, the design of starter cultures for reduced NaCl cheeses is recommended.

Plasmin activity in UHT milk: relationship between proteolysis, age gelation, and bitterness.

Plasmin, the major indigenous protease in milk, is linked to quality defects in dairy products. The specificity of plasmin on caseins has previously been studied using purified caseins and in the indigenous peptide profile of milk. We investigated the specificity and proteolytic pathway of plasmin in directly heated UHT milk (>150 °C for <0.2 s) during 14 weeks of storage at 20 °C in relation to age gelation and bitter peptides. Sixty-six peptides from αS- and ß-caseins could be attributed to plasmin activity during the storage period, of which 23 were potentially bitter. Plasmin exhibited the highest affinity for the hydrophilic regions in the caseins that most probably were exposed to the serum phase and the least affinity for hydrophobic or phosphorylated regions. The proteolytic pattern observed suggests that plasmin destabilizes the casein micelle by hydrolyzing casein-casein and casein-calcium phosphate interaction sites, which may subsequently cause age gelation in UHT milk.

Antioxidant properties of green tea extract protect reduced fat soft cheese against oxidation induced by light exposure.

The effect of two different antioxidants, EDTA and green tea extract (GTE), used individually or in combination, on the light-induced oxidation of reduced fat soft cheeses (0.2 and 6% fat) was investigated. In samples with 0.2% fat, lipid hydroperoxides as primary lipid oxidation products were not detected, but their interference was suggested from the formation of secondary lipid oxidation products such as hexanal and heptanal. The occurrence of these oxidation markers was inhibited by spiking with 50 ppm EDTA or 750 ppm GTE, or a combination of the two prior to irradiation. In contrast, addition of 50 ppm EDTA to samples with 6% fat was ineffective, but 750 ppm GTE (alone or in combination with EDTA) strongly reduced levels of hexanal and heptanal. Accumulation of primary lipid hydroperoxides was not affected by GTE, hence antioxidative activity was ascribed to scavenging of hexanal and heptanal precursors. These radical intermediates result from hydroperoxide disintegration, and subsequent scavenging by GTE, which acts as a radical sink, corroborates the intense signal observed by electron paramagnetic resonance (EPR) spectroscopy.

Deposition of carotenoids in egg yolk by short-term supplement of coloured carrot (Daucus carota) varieties as forage material for egg-laying hens.

BACKGROUND: Supplying egg-laying hens with different forage materials may influence egg production and quality. The aim of this study was to examine the short-term effects of standard feed plus 70 g day(-1) per hen of three coloured carrot varieties (orange, yellow and purple) as forage material in comparison with a standard feed control on egg production, egg yolk colour and deposition of carotenoids in the yolk. RESULTS: Carrot supplementation reduced feed intakes significantly, but not on a dry matter basis. Orange carrot treatment significantly reduced egg mass production, whereas yellow and purple carrot treatments did not differ from the control. Egg and yolk weights of all carrot-supplemented treatments were significantly lower than those of the control, but yolk percentages were similar. Yolk redness increased significantly in the order control < yellow < orange < purple. A similar trend was seen for yolk yellowness, but yellow and orange carrots reached the same level. Yolk colour and carotenoid contents correlated positively and significantly. In particular, purple carrot treatment increased the yolk content of lutein (>1.5-fold) and beta-carotene (>100-fold) compared with the control. CONCLUSION: Supplementing the feed of egg-laying hens with coloured carrots efficiently increased yolk colour parameters and carotenoid contents, which gives opportunities for improved nutritional value of eggs from forage material-supplemented hens.

Effect of grass-clover forage and whole-wheat feeding on the sensory quality of eggs.

BACKGROUND: A sensory panel evaluated the sensory profile of eggs from hens from three experimental systems: (1) an indoor system x normal layer diet (InL), (2) a grass-clover forage system x normal layer diet (GrL), and (3) a grass-clover forage system x whole wheat and oyster shells (GrW). RESULTS: The taste of the albumen was significantly more 'watery' and the yolks a darker yellow/orange in the eggs from the GrL and GrW groups. The yolk was darkest from the GrW group. The yolks from the InL and GrW groups had a significantly more 'fresh', less 'animal', 'cardboard', and 'intense' aroma than the GrL group. The taste of the yolks from the InL and GrW groups was significantly more 'fresh' and less 'cardboard'-like compared to the GrL group. The yolks tasted significantly less 'sulfurous' in the GrW group than in the GrL group. CONCLUSION: The combination of a high feed intake from a grass-clover pasture and the type of feed allocated is an important factor in relation to the sensory quality of eggs. Thus, a less favourable sensory profile of eggs was found from hens on a grass-clover pasture and fed a normal layer diet.