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Experimentally determined vitamin degradation kinetic parameters are vital input for process optimization, e.g., of nutrient rich beverages such as milk and fruit juices. The standard method of obtaining these parameters is to subject a test-tube of the nutrient (in its food matrix) to a water bath at elevated temperatures and measure remaining concentration as a function of time. It is well known that accurate kinetic fitting requires measuring the temperature inside the test tube as a function of time (due to lag it is not identical to the water-bath temperature), and this is included in contemporary studies. However, the importance of axial temperature gradients for valid kinetic parameter estimations has not been discussed to the same extent. This contribution combines experiments and physical modelling to show how an axial temperature gradient evolves in a test tube (R = 13 mm, Z = 75 mm), due to free convection currents (T∞ – T0 = 57–71 °C). Consequently, the test tube temperature is not uniform in such an experiment. Thus, the positioning of the thermocouple in a test-tube-in-water bath experiment is essential for ensuring a valid estimation of the kinetic parameters. Simulations suggest that positioning it half-way between top and bottom is a suitable best practice.

During whey powder production, the feed is subjected to several heat treatments which can cause lactosylation of proteins. In this study, lactosylation of whey proteins was evaluated in spray-dried powders before and after storage by varying the native protein fraction as well as the serum protein/lactose ratio in the powders. The lactosylation of native α-lactalbumin and β-lactoglobulin in the powders before storage was not affected to a large extent by the protein denaturation or if the feed had been heat treated in a high or low lactose environment. After storage (relative humidity of 23.5%, 30 °C, 25 days), the kinetic of lactosylation tended to increase with increasing native protein fraction and bulk protein content in the powders. An explanation could be that proteins dissolved in the lactose glassy structure might have a lower reactivity, while proteins present in the protein glassy structure with dissolved lactose may display higher lactosylation reactivity. (Less)

Spray-dried powders were produced from milk serum protein concentrate and lactose in varying ratios, and the rehydration characteristics of the powders were evaluated. The dissolution rate was estimated with a flow-cell based technique, and the external and internal distribution of the powder components were evaluated with X-ray photoelectron spectroscopy and confocal Raman microscopy, respectively. The surface of the powder particles is more or less covered by a thin protein layer. A phase segregation between protein and lactose is observed in the interior of the particle resulting in a protein rich layer in the vicinity of the surface. However, the protein layer in the vicinity of the particle surface tends to become thinner as the bulk protein concentration increases in the powders (from 10 to 60% w/w). The time for the spontaneous imbibition to occur show a linear correlation with the protein surface coverage. The dissolution rate of powders containing 0.1% w/w protein is around 60 times faster than for a powder containing 1% w/w protein but the dissolution rate of powders containing 1% and 100% w/w differ only by a factor of 2. Thus, it is suggested that the outer protein layer becomes denser at the interface as the protein content increases in the powders, thereby causing poorer rehydration characteristics of the powders (especially for low protein concentrations 0.1–1% w/w). This insight has relevance for the formulation of whey protein powders with improved rehydration characteristics.

A rigid mineral-based deposit, termed ultra-high temperature (UHT) fouling, is formed on heat exchanger surfaces during milk processing at 140 °C. The removal of this type of fouling is for the first time assessed using a laser triangulation sensor and a camera, to measure changes in the thickness and to visualize structural changes in the fouling in situ. The process was monitored during both the alkali and acid cleaning steps. There was no global swelling of the fouling layer during alkali cleaning under the investigated conditions. However, significant degradation of the protein network was observed, which affected the acid cleaning step and the efficiency of the cleaning process. We conclude that treatment with alkali is required to facilitate the removal of deposits with high mineral content during the acid cleaning step. The results have implications for optimizing the cleaning process so as to minimize energy expenditure, while ensuring efficient heat transfer and maintaining product quality.

Milk that does not coagulate after rennet addition, also called noncoagulating (NC) milk, is unwanted in cheese production due to prolonged processing time. Amounts of whey and casein proteins, genetic variants, as well as posttranslational modifications (PTM) of proteins are all contributing factors in rennet-induced coagulation of milk. In this study, we conducted a wide-ranging investigation of milk proteins in milk samples from 616 Swedish Red dairy cattle using liquid chromatography-high resolution mass spectrometry. Relative concentration of proteins, genetic variants, and PTM were compared between NC milk and coagulating milk. The PTM investigated were phosphorylation of caseins and glycosylation of κ-casein. Several genetic variants and PTM were found, including rare phosphorylation variants of the αS-caseins. Genetic variants were found to effect the expressed amount of different proteins. Further, the effect of protein amounts and PTM on a binary NC milk trait was modeled using a generalized linear model. The model showed that NC milk significantly correlated with higher relative concentrations of α-lactalbumin and β-casein and lower relative concentrations of β-lactoglobulin and κ-casein. Regarding PTM of caseins, an effect on NC milk from a lower relative concentration of αS1-casein with 8 phosphate groups were found, even though an effect from total relative concentration of αS1-casein was not found. This study has provided insights into protein variants and PTM important for NC milk to improve this undesirable property.

The rennet-induced coagulation ability of milk is important in cheese production. For Swedish Red Dairy Cattle (RDC), this ability is reduced because of a high prevalence of noncoagulating (NC) milk. In this study, we simultaneously combined genetic parameters for NC milk, milk coagulation properties, milk composition, physical traits, and milk protein composition. Our aim was to estimate heritability and genetic and phenotypic correlations for NC milk and 24 traits (milk coagulation properties, milk composition, physical traits, and milk protein composition). Phenotypes and ~7,000 SNP genotypes were available for all 600 Swedish RDC. The genotypes were imputed from ~7,000 SNP to 50,000 SNP. Variance components and genetic parameters were estimated with an animal model. In Swedish RDC, a moderate heritability estimate of 0.28 was found for NC milk. For the other 24 traits, heritability estimates ranged from 0.12 to 0.77 (standard errors from 0.08 to 0.18). A total of 300 phenotypic and genetic correlations were estimated. For phenotypic and genetic correlations, 172 and 95 were significant, respectively. In general, most traits showing significant genetic correlations also showed significant phenotypic correlations. In this study, phenotypic and genetic correlations with NC milk suggest that many correlations between traits exist, making it difficult to predict the real consequences on the composition of milk, if selective breeding is applied on NC milk. We speculate that some of these consequences may lead to changes in the composition of milk, most likely affecting its physical and organoleptic properties. However, our results suggest that κ-casein could be used as an indicator trait to predict the occurrence of NC milk at the herd level.

This study investigated milk serum protein concentrate/lactose systems with varying ratios and how the morphology of the spray-dried particles of these systems could be described by the surface properties of the feed as well as the protein surface coverage of the particles. An extrapolation of the surface pressure of the feed to 0.3 s, the approximate time for molecular diffusion in an atomised droplet in the spray-dryer, showed a relationship with the particle morphology. At low protein concentrations (

Complete wetting is crucial for efficient recombination of powders. On the powder bed scale, wetting is governed solely by capillary forces and the resistance to flow, i.e. spontaneous imbibition. Slow or incomplete imbibition of the powder bed may lead to gelling of the liquid front, which will stop the recombination process and cause the formation of lumps, which is usually undesirable. In this study, the spontaneous imbibition of powder beds consisting of spray-dried dairy powders with diverse morphologies has been investigated. Uniform radial spreading of the imbibition front was seen in all the beds, but a large amount of air was trapped in the inter-particle free space in the imbibed volume. A positive correlation was found between bed porosity and the fraction of air trapped in the bed after imbibition. Since the amount of trapped air was calculated as a fraction of the porosity of the dry bed, this relation was unexpected. The large fraction of air trapped in the bed and the uniform radial spreading of the imbibition front indicate considerable heterogeneity of bed on the microscopic scale, but a homogenous structure on the macroscopic scale. Possible explanations of the large fraction of trapped air are the presence of local dead-end structures in the bed and film flow of the imbibing liquid.

Milk oligosaccharides are of high interest due to their bioactive properties. This study is the first to characterise milk oligosaccharides from native North European cattle breeds, as represented by 80 milk samples collected from eight native breeds originated from Norway (Norwegian Doela cattle and Norwegian Telemark cattle), Sweden (Swedish Mountain cattle), Denmark (Danish Red anno 1970), Iceland (Icelandic cattle), Lithuania (native Lithuanian Black and White) and Finland (Western Finncattle and Eastern Finncattle). Using high-performance liquid-chromatography chip/quadrupole time-of-flight mass-spectrometry, 18 unique monosaccharide compositions and a multitude of isomers were identified. No N-glycolylneuraminic acid was identified among these breeds. Western Finncattle milk was most abundant in neutral, acidic and fucosylated oligosaccharides. Further, Eastern Finncattle milk was significantly higher in acidic oligosaccharides and Icelandic cattle milk significantly higher in fucosylated oligosaccharides, compared to the mean. This study highlights specific native breeds of particular interest for future exploitation of milk oligosaccharides and breeding strategies.

To keep their functional properties, it is crucial that protein aggregates maintain their structure after spray drying and that the powders can be fully rehydrated. In this study, microgels and fractal aggregates were prepared by heating a mixture of milk serum protein concentrate and lactose (40/60; %, w/w) at 85 °C for 15 min by varying the pH. Various fractions of native proteins were added to the systems prior to spray drying. This study showed that microgels and fractal aggregates kept their structure after spray drying and reconstitution. The particle morphology could be correlated to the stiffness of the interface of the feed droplet. The forced imbibition rate showed a negative correlation with increasing amount of aggregated proteins in the powders that seems to be a result of denatured/aggregated proteins present at the surface. These findings are of importance for the formulation of spray-dried powders with improved rehydration characteristics.

Whey powder rehydration is expected to be closely linked to both denaturation and lactosylation of the proteins. This study investigated the relation between the forced imbibition rate of spray-dried milk serum protein/lactose powders and the particle morphology and how it is related to the insoluble and lactosylated protein fraction, respectively. Despite extensive variation in protein denaturation, aggregation and lactosylation, only comparably small effects on the forced imbibition rate of the powders and the particle morphology could be observed. A possible explanation for this rather limited effect on the rehydration properties and particle morphology might be that the surface composition of the powder particles is rather similar and dominated by native proteins. These insights have relevance for the formulation of whey powders with improved rehydration properties.

The objective of this study was to investigate the influence of native milk fat globule size on the aeration of high fat dairy products with regard to maximum firmness time, gas inclusion and foam stability. The results showed that whipping time to maximum firmness was inversely proportional to mean fat globule size for both unhomogenised and slightly homogenised (2 MPa) creams. Additionally, increasing native mean fat globule size of the creams resulted in increased overrun. No significant differences in serum drainage were found between creams with different native milk fat globule size. Furthermore, when creams with native large mean fat globules were homogenised, the results showed that the maximum firmness time was in accordance with the mean fat globule size of non-aggregated creams. In the present study, cream fractionation was achieved by creaming or in a cost effective and fast manner using a modified centrifugal separator.

The formation of lumps during industrial recombination of powder is a common problem in many industries. An important step in the recombination process is the wetting of the dry material. Only with an efficient wetting the individual powder particles will disperse and a complete dissolution will occur. In difference, a slow or incomplete wetting will not supply liquid to all portions of the powder, resulting in the formation of lumps. The resistance to wetting can be described by a material's permeability. This parameter is often estimated using the Kozeny-Carman equation, which models the permeability of a porous medium (such as a bed of powder) based on the porosity and specific surface area of the medium. This model has proven valid for a bed of packed spheres, but due to the model assumptions the accuracy of the model when utilized on beds formed from more complex, spray dried powders can be questioned. In this study, the permeability of powder beds consisting of spray-dried diary powders when exposed to an air flux has been characterized, using a method based on air permeametry. The results show that the Kozeny-Carman model is insufficient to estimate the permeability of such beds. Furthermore, the measured porosity of the bed had no influence on the permeability constant. Based on the results of this study a correlation between morphological powder parameters and permeability was developed. This correlation indicates that a small surface weighted average diameter of the particles as well as a narrow width of the particle size distribution have a positive effect on the permeability of a bed. In addition, incorporation of the structural parameter “BedRatio”, defining the basis of the bed to consist of agglomerates or primary particles (fines), indicates that a bed consisting of mainly agglomerates has a slightly larger permeability.

© 2016 Elsevier Ltd. The fouling structure and composition is dependent on the product, but also on the heating process applied to it. The structure will have profound effect on the cleaning process and the down time in the production plant. Here, the structure of high temperature (137 °C) milk fouling has been investigated, which so far has not been sufficiently studied in a systematic way. This particular fouling has a high content of the mineral calcium phosphate and a relatively low concentration of protein. Wide angle X-ray diffraction (WAXD) reveals a crystalline structure of calcium phosphate in agreement to the chemical analysis of the bulk layer. Microscopic investigations visualize the heterogeneous structure and energy dispersive X-ray spectroscopy (EDX) shows a spatial variation of the elements through the radius of the sample.

Non-coagulating milk is unwanted in cheese production because it impairs chymosin-induced coagulation properties in bulk milk. Milk samples were analysed to investigate the coagulation process in non-coagulating milk for comparison with milk samples that have a high coagulation ability. This was done by measuring two peptides after chymosin cleavage, namely para-κ-casein and caseinomacropeptide, in all the milk samples using capillary electrophoresis. The results showed that cleavage of κ-casein into para-κ-casein and caseinomacropeptide occurred in both coagulation groups, suggesting that the aggregation stage is the cause of non-coagulating milk. Non-coagulating milk had lower content of para-κ-casein after chymosin addition compared with coagulating milk, which might be the result of a lower content of available κ-casein. Furthermore, this study indicates that there are different mechanisms that explain good coagulation ability and lack of coagulation ability of milk. The results can be useful in understanding the potential chemical mechanisms behind non-coagulating milk.

An increasing demand for dairy products with long shelf-life is foreseen. A limiting factor for maintaining high quality of these products is the activity of spoilage enzymes during storage. Lipase and protease activities in Swedish raw milk at farm and dairy level were investigated, analysing milk samples from three geographical regions and two seasons. Lipase activity in milk at farm level was affected by regional and seasonal variations, whereas at dairy level only season had an effect. Lipase activity was positively correlated with ionic calcium. For protease activity, no effect of either region or season was seen. Degradation products, e.g., free fatty acids, peptides and plasmin proteolysis products, varied differently between season and geographical origin at both farm and dairy level. The results indicate that lipase and protease activities are important for raw milk quality, while ionic calcium might be a future indicator for milk fat stability.

The objective of this study was to investigate how the properties of the serum phase, including ionic strength, protein content and specific milk salts, influence the whipping time of unhomogenised cream. This study showed that decreasing the protein content of the serum phase in cream resulted in shorter whipping time. However, addition of UF permeate to both deproteinated and proteinated creams indicated that other components than the protein fraction also influence whipping time of unhomogenised cream. Increasing the ionic strength by addition of NaCl, KCl and CaCl2 decreased whipping time, with the largest effect being found for CaCl2. A further increase in ionic strength by addition of NaCl and KCl resulted in an unfavourable effect on whipping time, whereas a pronounced effect, with shorter whipping times, was found for addition of CaCl2. (C) 2015 Elsevier Ltd. All rights reserved. (Less)

Butter is rich in saturated fat [saturated fatty acids (SFAs)] and can increase plasma low density lipoprotein (LDL) cholesterol, which is a major risk factor for cardiovascular disease. However, compared with other dairy foods, butter is low in milk fat globule membrane (MFGM) content, which encloses the fat. We hypothesized that different dairy foods may have distinct effects on plasma lipids because of a varying content of MFGM.We aimed to investigate whether the effects of milk fat on plasma lipids and cardiometabolic risk markers are modulated by the MFGM content.The study was an 8-wk, single-blind, randomized, controlled isocaloric trial with 2 parallel groups including overweight men and women (n = 57 randomly assigned). For the intervention, subjects consumed 40 g milk fat/d as either whipping cream (MFGM diet) or butter oil (control diet). Intervention foods were matched for total fat, protein, carbohydrates, and calcium. Subjects were discouraged from consuming any other dairy products during the study. Plasma markers of cholesterol absorption and hepatic cholesterol metabolism were assessed together with global gene-expression analyses in peripheral blood mononuclear cells.As expected, the control diet increased plasma lipids, whereas the MFGM diet did not [total cholesterol (±SD): +0.30 ± 0.49 compared with -0.04 ± 0.49 mmol/L, respectively (P = 0.024); LDL cholesterol: +0.36 ± 0.50 compared with +0.04 ± 0.36 mmol/L, respectively (P = 0.024); apolipoprotein B:apolipoprotein A-I ratio: +0.03 ± 0.09 compared with -0.05 ± 0.10 mmol/L, respectively (P = 0.007); and non-HDL cholesterol: +0.24 ± 0.49 compared with -0.14 ± 0.51 mmol/L, respectively (P = 0.013)]. HDL-cholesterol, triglyceride, sitosterol, lathosterol, campesterol, and proprotein convertase subtilisin/kexin type 9 plasma concentrations and fatty acid compositions did not differ between groups. Nineteen genes were differentially regulated between groups, and these genes were mostly correlated with lipid changes.In contrast to milk fat without MFGM, milk fat enclosed by MFGM does not impair the lipoprotein profile. The mechanism is not clear although suppressed gene expression by MFGM correlated inversely with plasma lipids. The food matrix should be considered when evaluating cardiovascular aspects of different dairy foods. This trial was registered at clinicaltrials.gov as NCT01767077.

Effects of dynamic ultra-high pressure homogenisation (UHPH) on the micro-and macrostructure of whey protein-depleted milk concentrate solutions with different mineral concentrations (5.0, 7.5, 10.0%, w/w), pH (5.8, 6.2, 6.6) and amount of added sodium chloride (0, 1.5, 3.0%, w/w) were investigated. Significantly increased viscosity was observed in samples at pH 5.8 without added NaCl and with UHPH treatment of 300 MPa and this effect was more pronounced at low mineral level. In this sample, irrespective of mineral content, an increased particle size was also found. The lowest water mobility was observed in samples with low mineral level, low pH and no added NaCl. The results indicate that two-dimensional longitudinal-transverse relaxation nuclear magnetic resonance (NMR) measurements can contribute to a better understanding of microstructure formation in whey protein-depleted milk concentrate. Application H-1 and P-31 NMR spectroscopy indicated that UHPH treatment only affected protein aggregation and not casein micelle structure. (C) 2014 Elsevier Ltd. All rights reserved. (Less)

Milk casein concentrates with different casein-to-protein ratios (0.82 and 0.93), pH (5.8, 6.2 and 6.7) and level of applied ultra-high pressure homogenisation (UHPH) (0, 150 and 300 MPa) were studied. Samples with a casein-to-protein ratio of 0.93 at pH 5.8 and subjected to UHPH treatment at 300 MPa had the highest apparent viscosity and particle size followed by similar samples with a casein-to-protein ratio of 0.82. Water mobility was reduced at higher casein-to-protein ratio and with higher pH, while increased water mobility was observed in samples with a casein-to-protein ratio of 0.82 at pH 5.8 subjected to UHPH treatment at 300 MPa. The H-1 and P-31 NIVIR spectra revealed minor structural effects of adding casino-phospho-peptides and changing pH, but no effect of UHPH treatment, which indicate that UHPH treatment did not induce measureable structural changes in the individual proteins but rather induced changes in the tertiary structure or degree of polymerisation. Industrial relevance: Milk depleted from whey protein is advantageous to use in cheese production since it can be further processed without detrimental denaturation of whey protein which will result in reduced maturation of cheese. In combination with pH control and ultra-high pressure homogenisation of the milk casein concentrate structural changes in the protein can be obtained and new interesting milk based products can be developed. (C) 2014 Elsevier Ltd. All rights reserved. (Less)

Chymosin-induced gelation properties of the milk of around 400 Swedish Red dairy cows was investigated with the aim of identifying genetic protein variants that influence chymosin-induced gelation for optimisation of cheese milk through breeding practices. The variation in chymosin-induced gelation properties was shown to be large in the milk of Swedish Red and there was a high frequency of cows producing non-coagulating milk, 18%. The present study showed that the common composite beta-kappa-casein genotypes A(1)A(2)/AE and A(2)A(2)/AA were associated with both poor gelation properties and non-coagulation in the milk of Swedish Red. The present study suggests that if the frequencies of composite genotypes A(1)A(2)/AE and A(2)A(2)/AA were decreased in the Swedish Red population in favour for A(1)A(1)/AA and A(1)A(1)/AE, this could have a positive effect on the rennetability of the milk. (C) 2014 Elsevier Ltd. All rights reserved.

Changes occurring in directly heated UHT milk were studied during storage at 5, 22, 30 and 40 °C. Industrially produced UHT milk samples were analysed for changes in enzymatic activity, protein modification, destabilisation of casein micelles and relocation of milk proteins in relation to sedimentation and gel formation. Sedimentation occurred at all temperatures, and the protein composition of the sediments reflected the composition of its liquid phase; however, there was no α-lactalbumin, β-lactoglobulin or κ-casein present in sediments. Tendrils composed of β-lactoglobulin and κ-casein were seen on casein micelles after UHT treatment and grew in length prior to gelation. High degrees of lactosylation of proteins and peptides were clearly correlated with the absence of gelation and long tendrils. Gelled samples showed complete hydrolysis of intact β-casein, and limited lactosylation of β-lactoglobulin and κ-casein.

Recent studies have reported a very high frequency of noncoagulating milk in Swedish Red cows. The underlying factors are not fully understood. In this study, we explored rennet-induced coagulation properties and relative protein profiles in milk from native Swedish Mountain and Swedish Red Polled cows and compared them with a subset of noncoagulating (NC) and well-coagulating (WC) milk samples from modern Swedish Red cows. The native breeds displayed a very low prevalence of NC milk and superior milk coagulation properties compared with Swedish Red cows. The predominant variants in both native breeds were αS1-casein (αS1-CN) B, β-CN A2 and β-lactoglobulin (β-LG) B. For κ-CN, the B variant was predominant in the Swedish Mountain cows, whereas the A variant was the most frequent in the Swedish Red Polled. The native breeds displayed similar protein composition, but varied in content of αS1-CN with 9 phosphorylated serines (9P) form. Within the Swedish Mountain cows, we observed a strong inverse correlation between the relative concentration of κ-CN and micelle size and a positive correlation between ionic calcium and gel firmness. For comparison, we investigated a subset of 29 NC and 28 WC milk samples, representing the extremes with regard to coagulation properties based on an initial screening of 395 Swedish Red cows. In Swedish Red, NC milk properties were found to be related to higher frequencies of β-CN A2, κ-CN E and A variants, as well as β-LG B, and the predominant composite genotype of β- and κ-CN in the NC group was A2A2/AA. Generally, the A2A2/AA composite genotype was related to lower relative concentrations of κ-CN isoforms and higher relative concentrations of αS1-, αS2-, and β-CN. Compared with the group of WC milk samples, NC milk contained a higher fraction of αS2-CN and α-lactalbumin (α-LA) but a lower fraction of αS1-CN 9P. In conclusion, milk from native Swedish breeds has good characteristics for cheese milk, which could be exploited in niche dairy products. In milk from Swedish Mountain cows, levels of ionic calcium seemed to be more important for rennet-induced gel firmness than variation in the relative protein profile. In Swedish Red, lower protein content as well as higher fraction of αS2-CN and lower fraction of αS1-CN 9P were related to NC milk. Further, a decrease in the frequency of the composite β-κ-CN genotype A2A2/AA through selective breeding could have a positive effect on milk coagulation properties.

A spectrophotometric assay for plasmin and plasminogen-derived activity in dairy products was optimised and extended to determine plasmin and plasminogen-derived activity in turbid samples of dairy products. The method was validated by assessing reproducibility, repeatability, level of detection and recovery of plasmin activity in different sample matrices. Plasmin activity in raw milk was not affected by skimming, but decreased by 30% in pasteurised and homogenised whole milk, leading to an underestimation of plasmin activity. The effects of dissociation of plasmin and caseins by epsilon-aminocaproic acid (EACA) plus NaCl on the plasmin activity were investigated. Comparison of pasteurised milk with a micellar casein solution showed that the dissociation of plasmin and caseins on adding EACA and NaCl decreases interference by caseins, but increases inhibition of plasmin with serum-based inhibitory components. The level of detection and repeatability of this method for plasmin activity analysis were improved compared with previous spectrophotometric assays. (C) 2014 Elsevier Ltd. All rights reserved. (Less)

The formation of lumps during large scale wetting of spray dried powders is a common problem in the industry. The result is a large energy demand if an efficient mixing is to be achieved at the industrial scale. If more knowledge regarding the phenomena governing lump formation is obtained, the unit operation could be optimized and made more efficient. As a part of increasing the understanding of lump formation, the wetting of the porous media that is a powder bed needs to be understood. This wetting relates to the structure and void space (porosity) of the powder bed, which is why a relation between powder bed porosity and readily obtainable powder particle morphology parameters is sought. In this study a thorough investigation of the morphology of five different spray dried milk powders, with arbitrary particle shapes, was performed using manual image analysis of scanning electron microscope (SEM) images as well as automated methods based on light scattering and projected image analysis. The results of the automated methods, when utilized on the complexly shaped samples, were validated by a comparison to the SEM image analysis. This enabled the development of a relation between powder morphology and powder bed porosity based on data from automated methods. The results indicate that automated methods based on light scattering underestimate the particle sizes of arbitrary shaped particles. Methods based on projected image analysis, however, show a better agreement with SEM image analysis. The developed relation between powder bed porosity and powder particle morphology is based on defining the bed as having either an "Agglomerates in fines", "Bicontinuous" or "Fines in agglomerates" structure. The structure of each powder is determined based on calculation of a "BedRatio" which is a function of the phi(agglomerate) and phi(fines) both shown to be readily obtainable by automated methods. The calculated porosities of the powders investigated in this study show a good agreement with experimental measurements and are considered to correspond well to earlier literature. It is, however, clear from the a experiments that care needs to be taken during bulk density measurement of spray dried powders so that no intra particle void space is taken into consideration in the calculations. (C) 2013 Elsevier B.V. All rights reserved. (Less)

Ultra-high temperature (UHT) treatment of milk forms a deposit or fouling in the processing equipment that is mineral-based with an enclosed protein network. This study addresses the fundamental mechanisms that control the removal of this deposit. For this purpose, the structural and compositional changes during the cleaning process have been studied. The structure analysis was performed with scanning electron microscopy (SEM) and confocal laser scanning microscopy (CLSM) on samples that were quenched at different stages of the cleaning process. It was found for acid cleaning that the mineral content is rapidly decreasing in the fouling layer as the cleaning continues, but there is still an intact protein structure with the similar thickness as the original fouling. For alkali cleaning, part of the protein structure was subsequently removed from the outside towards the stain-less steel as a function of time, while the mineral structure was mostly remaining. The break-up of the organic network structure, which likely involves depolymerization of protein aggregates, were found to control the cleaning efficiency. The weakening of the protein network facilitates the removal of the UHT fouling layer during the acid cleaning step and allow for an efficient cleaning cycle. The chemical reactions that occur within the fouling layer between the hydroxyl ions and the protein network was modeled according to a depolymerization reaction and a mechanistic model of the cleaning process is presented.

The formation of lumps is a common problem during powder reconstitution in food industry. If more knowledge regarding the phenomena controlling the process could be obtained, the process of industrial powder recombination could be optimized. This demands investigation of the process at three different levels: the scale of individual powder particles, the scale of powder beds, and the scale of industrial recombination. If knowledge from all three scales could be combined, a model of the entire process could be constructed. In this study, we present a new flow-cell-based method for investigating the dissolution rate of individual powder particles in controlled liquid flow. The flow cell allows for alteration of the flow velocity and only subjects the investigated powder particles to mild stress before the analysis. The dissolution process in the cell is recorded, and data are obtained by image analysis. Sodium caseinate has been used as a model substance in these experiments. The results show that the particle dissolution rate is a function of the flow velocity of the liquid, and the results correspond to earlier established methods. The investigation of individual particles shows a variation in dissolution rate between particles resulting from the diverse particle morphology from an industrial batch of powder. The results indicate that the dissolution interface concentration of dissolving sodium caseinate particles corresponds well to the overlap concentration of the specie.

The composition of milk fat from dairy cows is related to both genetic and environmental factors. Here, the effect of feed and herd was examined in 3 Scandinavian breeds, namely Danish Holstein-Friesian (DH), Danish Jersey (DJ), and Swedish Red (SR). In total, milk samples from 1,298 cows kept in indoor housing systems were collected from 61 conventional dairy herds in Denmark and Sweden. The fatty acid (FA) composition of milk was determined by gas chromatography and the content of α-tocopherol by HPLC. Based on the 17 individual FA determined, distinct FA profiles were observed for all breeds using univariate and multivariate statistics. The DJ cows were characterized by higher levels of saturated short-chain FA; in contrast, DH cows had higher content of unsaturated C18 FA, whereas higher levels of primarily C14:0, C14:1, C18:1 cis-9, and C18:3n-3 were evident in SR cows. This variation in milk fat composition across breeds was further reflected in different desaturase indices, which were generally higher in SR cows. In addition, α-tocopherol differed significantly among breeds, with DJ cows having the highest content. Herd-specific feeding plans were collected, and different feed items were separated into 4 broad feed categories, including grass products, maize silage, grain, and concentrate. The pronounced differences in overall feed composition among breeds were, to a large extent, due to regional differences between countries, with SR receiving higher levels of grain and grass silage compared with the Danish breeds. Within breeds, differences in feeding regimens among herds were furthermore higher in SR. Significant correlations between feed category and individual FA were observed in all breeds. Furthermore, variance components were estimated and used to determine the proportion of phenotypic variation that could be explained by herd. The herd effect for individual FA was generally lower for DH compared with the 2 other breeds. In addition, very low herd effects were shown for C14:1 and C16:1 in all breeds, suggesting that the content of these FA is mainly genetically regulated.

Treatment of Caco-2 cells with the peptide lactoferricin(4-14), results in reduction of the growth rate by prolongation of the S phase of the cell cycle. Lactoferricin(1-25) is formed in the gut by cleavage from lactoferrin and the bioactive amino acids are found within lactoferricin(4-14). Our hypothesis is that the reduction of the rate of S phase progression may result in increased DNA repair. To test this hypothesis, Caco-2 cells were subjected to UV light that caused DNA lesions and then the cells were grown in the absence or presence of 2.0 μM lactoferricin(4-14). Evaluation of DNA strand breaks using the comet assay showed that lactoferricin(4-14) treatment indeed resulted in a reduction of comets showing damaged DNA. In the search for a mechanism, we have investigated the levels of several proteins involved in cell cycle regulation, DNA replication, and apoptosis using Western blot. Lactoferricin(4-14) treatment resulted in an increased expression of flap endonuclease-1 pointing to increased DNA synthesis activity. Lactoferricin(4-14) treatment decreased the expression of the proapoptotic protein B-cell lymphoma 2-associated X protein (or Bax), indicating decreased cell death. As we have found previously, lactoferricin(4-14) treatment reduced the expression of cyclin E involved in the G(1)/S transition. Immunofluorescence microscopy showed that a lower γ-H2AX expression in lactoferricin(4-14)-treated cells, pointing to more efficient DNA repair. Thus, altogether our data show that lactoferricin(4-14) treatment has beneficial effects.

In the native bovine casein micelle the calcium sensitive caseins (α(S1)-, α(S2)- and β-casein) sequester amorphous calcium phosphate in nanometer-sized clusters, whereas the calcium-insensitive κ-casein limits the growth of the micelle. In this paper, we further investigate the self-association of κ- and β-casein, which are two of the key proteins that control the substructure of the milk casein micelle, using neutron and light scattering techniques and cryogenic transmission electron microscopy. Results demonstrate that κ-casein can, apart from the known self-assembly, form amyloid-like fibrils already at temperatures of 25 °C when subject to agitation. This extended aggregation behavior of κ-casein is inhibited by β-casein, as reported by others. These findings have implications for the structure and stability of casein micelles. The neutron scattering data was used to gain information on the self-assembly structure of κ-casein. β-Casein shows similar self-association behavior as κ-casein, but unlike κ-casein, the self-association exhibits temperature dependence within the studied temperatures (6 and 25 °C). Here, we will discuss our extended study of the known self-assembly of casein in the context of the fibrillation of κ-casein.

Non-coagulating (NC) milk, defined as milk not coagulating within 40 min after rennet-addition, can have a negative influence on cheese production. Its prevalence is estimated at 18% in the Swedish Red (SR) cow population. Our study aimed at identifying genomic regions and causal variants associated with NC milk in SR cows, by doing a GWAS using 777k SNP genotypes and using imputed sequences to fine map the most promising genomic region. Phenotypes were available from 382 SR cows belonging to 21 herds in the south of Sweden, from which individual morning milk was sampled. NC milk was treated as a binary trait, receiving a score of one in case of non-coagulation within 40 min. For all 382 SR cows, 777k SNP genotypes were available as well as the combined genotypes of the genetic variants of αs1-β-κ-caseins. In addition, whole–genome sequences from the 1000 Bull Genome Consortium (Run 3) were available for 429 animals of 15 different breeds. From these sequences, 33 sequences belonged to SR and Finish Ayrshire bulls with a large impact in the SR cow population. Single-marker analyses were run in ASReml using an animal model. After fitting the casein loci, 14 associations at −Log10(P-value) > 6 identified a promising region located on BTA18. We imputed sequences to the 382 genotyped SR cows using Beagle 4 for half of BTA18, and ran a region-wide association study with imputed sequences. In a seven mega base-pairs region on BTA18, our strongest association with NC milk explained almost 34% of the genetic variation in NC milk. Since it is possible that multiple QTL are in strong LD in this region, 59 haplotypes were built, genetically differentiated by means of a phylogenetic tree, and tested in phenotype-genotype association studies. Haplotype analyses support the existence of one QTL underlying NC milk in SR cows. A candidate gene of interest is the VPS35 gene, for which one of our strongest association is an intron SNP in this gene. The VPS35 gene belongs to the mammary gene sets of pre-parturient and of lactating cows.

Genomic selection is a new technology in which selection decisions are based on direct genomic values (DGVs) or genomic enhanced breeding values (GEBVs). The objective of this study was to evaluate the relations between DGVs and several milk traits important for both the nutritional value and processability of milk. This is a new approach and can be used to increase the knowledge on how genomic selection can be used in practice. Morning milk samples from Swedish Holstein cows were analyzed for milk composition and technological properties. DGVs were received for each cow for milk, protein and fat yield, milk index, udder health, Nordic total merit and a quota was calculated between fat and milk yield as well as protein and milk yield. The results show that linear correlations exist (PP=0·002–0·097), fat (P=0·024–0·055) and mineral profiles (P=0·001–0·099) as well as for cheese characteristics (P=0·004–0·065), thus making it possible to obtain detailed information on milk traits that are not registered in the milk recording scheme. Hence, genomic selection will be an efficient tool for breeding and dairy industry to select cows early in life for targeted milk production.

Cheese production has increased worldwide during the last decade and is expected to increase within the coming decade as well. Despite this, the relations between cow genetics and cheese characteristics are not fully known. The aim of this study was to determine if polymorphisms in the leptin (LEP), leptin receptor (LEPR), and acyl-coenzyme A:diacylglycerol acyltransferase 1 (DGAT1) genes as well as genetic variants of β-casein (β-CN), κ-CN, and β-lactoglobulin (β-LG) affect technological properties important for cheese production and, hence, could act as genetic makers for cheese quality. Individual milk samples from the Swedish Red and the Swedish Holstein breeds were analyzed for sizes of CN micelles and fat globules as well as rennet-induced gel strength, gelation time, and yield stress. Model cheeses were produced to study yield, hardness, and pH of the cheeses. The A1457G, A252T, A59V, and C963T single nucleotide polymorphisms (SNP) were analyzed on the LEP gene, the T945M SNP on the LEPR gene, and the Nt984+8(A-G) SNP on the DGAT1 gene. In addition, genetic variants of β-CN, κ-CN, and β-LG were determined. The results indicate that technological properties were influenced by the LEPR(T945M) polymorphism, which had an association with gel strength, yield stress, and cheese hardness (T > C). However, also LEP(A252T) was shown to affect gel strength (T > A), whereas the LEP(A59V) had an effect on fat globule size (T > C). For the milk protein genes, favorable effects were found for the A and B variants of β-LG and κ-CN, respectively, on gel strength, gelation time, and yield stress. In addition, the B variant of κ-CN was shown to be associated with smaller CN micelles than the A variant. Thus, the results demonstrate potential genetic markers for cheese characteristics. However, milk composition traits also affected the obtained results, thus making it necessary to thoroughly assess the different aspects regarding the influence of gene effects on cheese characteristics before directly selecting for certain alleles or genetic variants to improve the processing and quality of cheese.

Casein (CN) micelles are naturally occurring colloidal protein aggregates present in a dispersed state in milk. In this paper we aim to obtain a detailed description of physicochemical properties of CN micelles over the entire size distribution using asymmetrical flow field-flow fractionation (AsFlFFF) connected to multiangle light scattering (MALS) and refractive index (RI) detection. Conclusions are drawn on the colloidal level regarding shape and conformation by comparison with models of colloidal particles. By using AsFlFFF-MALS-RI, it is concluded that the CN micelles are highly polydisperse with an average rms radius and hydrodynamic radius of 177 and 116 nm, respectively. The results show that the majority of CN micelles have a spherical shape, whereas a low concentration exists of larger and elongated aggregates. By comparison with models of aggregates of colloidal particles, the aggregates are shown to be anisotropic, e.g., aggregating linearly (threadlike) or in a sheet, rather than forming randomly spherical clusters. The results show that the characterization of colloidal dispersions with AsFlFFF-MALS-RI and the comparison with theoretical models are of a general character and, thus, of fundamental importance for colloidal dispersions.

The economic output of the dairy industry is to a great extent dependent on the processing of milk into other milk-based products such as cheese. The yield and quality of cheese are dependent on both the composition and technological properties of milk. The objective of this study was to evaluate the importance and effects of casein (CN) micelle size and milk composition on milk gelation characteristics in order to evaluate the possibilities for enhancing gelation properties through breeding. Milk was collected on 4 sampling occasions at the farm level in winter and summer from dairy cows with high genetic merit, classified as elite dairy cows, of the Swedish Red and Swedish Holstein breeds. Comparisons were made with milk from a Swedish Red herd, a Swedish Holstein herd, and a Swedish dairy processor. Properties of CN micelles, such as their native and rennet-induced CN micelle size and their zeta-potential, were analyzed by photon correlation spectroscopy, and rennet-induced gelation characteristics, including gel strength, gelation time, and frequency sweeps, were determined. Milk parameters of the protein, lipid, and carbohydrate profiles as well as minerals were used to obtain correlations with native CN micelle size and gelation characteristics. Milk pH and protein, CN, and lactose contents were found to affect milk gelation. Smaller native CN micelles were shown to form stronger gels when poorly coagulating milk was excluded from the correlation analysis. In addition, milk pH correlated positively, whereas Mg and K correlated negatively with native CN micellar size. The milk from the elite dairy cows was shown to have good gelation characteristics. Furthermore, genetic progress in relation to CN micelle size was found for these cows as a correlated response to selection for the Swedish breeding objective if optimizing for milk gelation characteristics. The results indicate that selection for smaller native CN micelles and lower milk pH through breeding would enhance gelation properties and may thus improve the initial step in the processing of cheese.

The initial stage of the Maillard reaction, protein lactosylation, occurs during heat treatment of milk and continues during subsequent storage. We compared the initial lactosylation as well as the rate of lactosylation of milk proteins during storage in UHT milk subjected to direct or indirect heat treatment using liquid chromatography (LC) coupled with electrospray injection mass spectrometry (ESI-MS). Furosine content was used as an overall marker to allow for a quantitative correlation of lactosylation measured by LC-ESI-MS in the UHT milks. Protein lactosylation increased during the storage period of 6months at 20 degrees C. Both the initial extent and the rate of lactosylation positively correlated with the number of lysine residues in the different proteins. An exponential or linear correlation with furosine concentration could be established for major and minor lactosylated proteins, respectively. (Less)

The production of fermented milk products has increased worldwide during the last decade and is expected to continue to increase during the coming decade. The quality of these products may be optimized through breeding practices; however, the relations between cow genetics and technological properties of acid milk gels are not fully known. Therefore, the aim of this study was to identify chromosomal regions affecting acid-induced coagulation properties and possible candidate genes. Skim milk samples from 377 Swedish Red cows were rheologically analyzed for acid-induced coagulation properties using low-amplitude oscillation measurements. The resulting traits, including gel strength, coagulation time, and yield stress, were used to conduct a genome-wide association study. Single nucleotide polymorphisms (SNP) were identified using the BovineHD SNPChip (Illumina Inc., San Diego, CA), resulting in almost 621,000 segregating markers. The genome was scanned for putative quantitative trait loci (QTL) regions, haplotypes based on highly associated SNP were inferred, and the additive genetic effects of haplotypes within each QTL region were analyzed using mixed models. A total of 8 genomic regions were identified, with large effects of the significant haplotype explaining between 4.8 and 9.8% of the phenotypic variance of the studied traits. One major QTL was identified to overlap between gel strength and yield stress, the QTL identified with the most significant SNP closest to the gene coding for κ-casein (CSN3). In addition, a chromosome-wide significant region affecting yield stress on BTA 11 was identified to be colocated with PAEP, coding for β-lactoglobulin. Furthermore, the coagulation properties of the genetic variants within the 2 genes were compared with the coagulation properties identified by the patterns of the haplotypes within the regions, and it was discovered that the haplotypes were more diverse and in one case slightly better at explaining the phenotypic variance. Besides these significant QTL comprising the 2 milk proteins, 3 additional genes are proposed as possible candidates, namely RAB22A, CDH13, and STAT1, and all have previously been found to be expressed in the mammary gland. To our knowledge, this is the first attempt to map QTL regions for acid-induced coagulation properties.

The somatic cell count (SCC) in bovine milk is an indicator of udder health and milk quality. Individual cell populations, i.e., macrophages, lymphocytes and polymorphonuclear neutrophils, were identified and their relationship to milk components such as acute-phase proteins (serum amyloid A (SAA) and α1-acid glycoprotein), lactic acid and lactoferrin (LF), as well as protein, fat and lactose, were studied in foremilk from separate udder quarters. The whey proteins and peptides were studied using two-dimensional gel electrophoresis, which was shown in the present study to be a useful method of assessing udder health and milk quality. The growth of a starter culture was evaluated using a conductance method as a simulation of fermentation processes relevant for cheese production. The investigation showed that LF, SAA, l-lactate and lactose are reliable indicators of milk quality on udder quarter level. The correlations between SCC, or the individual cell populations, and the potential indicators were highly significant.

Whey protein adsorption on stainless steel surfaces was investigated by in situ ellipsometry under well-defined flow conditions and protein solution residence times. The size distribution in solution of the protein aggregates formed under the same conditions was determined by dynamic light scattering. The adsorption was performed at 72 degrees C and 85 degrees C which is below and above the unfolding temperature of beta-lactoglobulin (75 degrees C). The effect of increasing the Reynolds number on the aggregation process was evident only for the longest protein solution residence time, when larger aggregates were produced. At the higher temperature, higher flow turbulence decreased the amount of protein adsorbed on the surface, which was attributed to shear-induced removal of protein from the surface. At both temperatures, longer protein solution residence times decreased both the adsorption kinetics and the amount of protein adsorbed. This could be related to the concentration of native protein available in solution. Based on the experimental results, a model of the formation of a protein monolayer and multilayers on solid surfaces was developed. The model takes into account the dependence of the adsorption process on both protein solution residence time and flow rate and predicts the evolution of adsorbed mass of protein per unit surface area with operating time. Surface-induced aggregation was identified as the main mechanism responsible for the formation of multilayers, based on experimental data. To reduce the amount of protein adsorbed/deposited on the surface an increase in protein solution residence time was found to be more effective than an increase in the Reynolds number. (c) 2005 Elsevier Ltd. All rights reserved. (Less)

The adsorption of whey proteins on modified and unmodified 316 2R stainless steel surfaces was studied by in situ ellipsometry under well-defined flow conditions. The effects of bulk properties, e.g. temperature, pH, protein concentration and flow rate as well as surface modification by ion implantation, sputtering, chemical vapour deposition and silica sol-gel deposition, were investigated. The surface properties were characterized in terms of wettability and roughness. The results suggest a surface-induced aggregation process given that (1) the adsorbed amount at high temperature was significantly larger than expected for a monolayer, and (2) the adsorption was lower with a larger number of aggregates in solution. The surface modifications affected the initial adsorption rate, i.e. the rate of forming the monolayer, which in turn influenced the surface induced aggregation and eventually the total amount deposited. Therefore, once the surface was fully covered with protein, the rate of deposition, which is controlled by protein-protein interactions, was less affected by the surface modifications. This work evidenced the potential of surface modification regarding the reduction of protein adsorption on steel surfaces.

Proteins and peptides in bovine milk and colostrum were separated by two-dimensional gel electrophoresis (2-DE). The proteins were separated in the first dimension by isoelectric focusing in the pH ranges 4-7 and 6-11, and in the second dimension by sodium dodecyl sulphate polyacrylamide gel electrophoresis on 12.5% T homogeneous gels. The peptides were separated in the pH range 3-10 in the first dimension, while 15% T homogeneous gels were used in the second dimension. 2-DE evaluation of samples with various somatic cell counts showed an increased number of peptides with increased cell count. 2-DE analysis of samples heated at 65 and 85 degreesC for 30 min showed a decrease in the number of proteins and peptides with increased temperature. Colostrum exhibited a peptide pattern in which the intensity and number of spots decreased with time postpartum. These results show that 2-DE is a useful tool to detect variations of proteins and peptides in milk. (Less)

Changes in the structure and chemistry of beta-lactoglobulin (beta-LG) play an important role in the processing and functionality of milk products. In model beta-LG systems, there is evidence that the aggregates of heated beta-LG are held together by a mixture of intermolecular non-covalent association and heat-induced non-native disulfide bonds. Although a number of non-native disulfide bonds have been identified, little is known about the initial inter- and intramolecular disulfide bond rearrangements that occur as a result of heating. These interchange reactions were explored by examining the products of heat treatment to determine the novel disulfide bonds that form in the heated beta-LG aggregates. The native protein and heat-induced aggregates were hydrolyzed by trypsin, and the resulting peptides, before and after reduction with dithiothreitol, were separated by high-performance liquid chromatography and their identities confirmed by electrospray ionization mass spectrometry. Comparisons of these peptide patterns showed that some of the Cys160 was in the reduced form in heated beta-LG aggregates, indicating that the Cys160-Cys66 disulfide bond had been broken during heating. This finding suggests that disulfide bond interchange reactions between beta-LG non-native monomers, or polymers, and other proteins could occur largely via Cys160.