How the presence of residual lipids in a yellow mealworm protein concentrate affects its foaming properties?

The use of whole and visible insects is poorly accepted in Western countries, and this remains a significant challenge for product development. However, using insect-based protein-rich ingredients, like protein concentrate, can improve levels of consumer approval. The residual lipid content in insect protein concentrates can influence their techno-functional properties. Our study therefore aimed to evaluate the impact of the residual lipid content on the protein structure and foaming properties of a mealworm protein concentrate. Our results showed that the protein content increased from 78.01 to 84.82 % after using chloroform-methanol for lipid removal. The particle size distribution shifted from a bimodal to a unimodal pattern, and the surface hydrophobicity decreased from 267.02 to 48.91 after completely removing lipids by chloroform-methanol, with no noticeable impact on the protein profile. The foaming capacity improved, resulting in the formation of a firm and fluffy foam with high stability over time. These results highlight the importance of controlling the residual lipid content in mealworm protein concentrates to enhance their techno-functional properties. The next steps will entail comprehensively characterizing the lipid profile and exploring the various mechanisms contributing to the techno-functional properties.

Influence of the processing on composition, protein structure and techno-functional properties of mealworm protein concentrates produced by isoelectric precipitation and ultrafiltration/diafiltration.

Edible insects represent a great alternative protein source but food neophobia remains the main barrier to consumption. However, the incorporation of insects as protein-rich ingredients, such as protein concentrates, could increase acceptance. In this study, two methods, isoelectric precipitation and ultrafiltration-diafiltration, were applied to produce mealworm protein concentrates, which were compared in terms of composition, protein structure and techno-functional properties. The results showed that the protein content of the isoelectric precipitation concentrate was higher than ultrafiltration-diafiltration (80 versus 72%) but ash (1.91 versus 3.82%) and soluble sugar (1.43 versus 8.22%) contents were lower. Moreover, the protein structure was affected by the processing method, where the ultrafiltration-diafiltration concentrate exhibited a higher surface hydrophobicity (493.5 versus 106.78 a.u) and a lower denaturation temperature (161.32 versus 181.44 °C). Finally, the ultrafiltration-diafiltration concentrate exhibited higher solubility (87 versus 41%) and emulsifying properties at pH 7 compared to the concentrate obtained by isoelectric precipitation.

Effect of high-hydrostatic pressure on the digestibility of egg yolk and granule.

The impact of high hydrostatic pressure (HHP) on protein digestibility of egg yolk and egg yolk granule was evaluated by static in vitro digestion using the standardized INFOGEST 2.0 method. The degree of hydrolysis (DH) and the phospholipid content were determined during digestion, and the protein and peptide profiles were characterized by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and reverse phase-high pressure liquid chromatography (RP-HPLC). The results showed that HHP induced protein aggregation in egg yolk and granule, mainly by disulfide bridges, which were not disrupted in the oral phase. Proteolysis during the gastric phase improved egg yolk and granule protein solubility, regardless of whether HHP was applied. However, the extent of the samples' digestibility was not affected, with DH values ranging from 15% to 20%. During the intestinal phase, the DH of egg yolk protein (∼40%) was higher than that of the granule (∼25%), probably due to the denser structure of the granule reducing the accessibility of intestinal enzymes. The DH, peptide, and protein profiles of control and HHP-treated egg yolk showed similar protein digestion behaviors for both gastric and intestinal phases. Among the different proteins, only the digestibility of ß-phosvitin in HHP-treated granule was enhanced. Consequently, applying HHP to granules represents an interesting process that improves the digestibility of phosvitin with the potential to generate bioactive phosvitin-derived phosphopeptides. PRACTICAL APPLICATION: High hydrostatic pressure, mainly used as a preservation process, did not impair the nutritional quality of the egg yolk and granule proteins but improved the susceptibility of phosvitin (protein contained in egg yolk) proteolysis to produce bioactive phosphopeptides. Consequently, applying HHP to granules represents an interesting process that improves the digestibility of phosvitin.

Effect of thermal and defatting treatments on the composition, protein profile and structure of house cricket (Acheta domesticus) protein extracts.

This study investigated the impact of blanching (100 °C, 40 s), defatting method (maceration, Soxhlet) and solvent polarity (hexane, ethanol) on the profile, structure and solubility of house cricket protein extracts. Blanching and Soxhlet using ethanol impacted the protein profile, with a lower content of myosin heavy chain and a higher abundance of low molecular weight proteins (<25 kDa). Moreover, ethanol induced aggregation of non-blanched cricket proteins, with a 13-72% reduction in protein recovery yield. The protein secondary structure of non-blanched extracts was also affected by ethanol with 18% more ß-sheets. Furthermore, blanching resulted in a lower protein surface hydrophobicity by a factor of 3 to 7, with no impact of solvent polarity. Finally, the solubility of protein extracts remained >75%, regardless of the blanching and defatting methods. These findings, combined with the evaluation of techno-functional properties, could be used for the development of cricket-based protein ingredients for food formulations.

Chronic Stress Exposure Alters the Gut Barrier: Sex-Specific Effects on Microbiota and Jejunum Tight Junctions.

Background: Major depressive disorder (MDD) is the leading cause of disability worldwide. Of individuals with MDD, 30% to 50% are unresponsive to common antidepressants, highlighting untapped causal biological mechanisms. Dysfunction in the microbiota-gut-brain axis has been implicated in MDD pathogenesis. Exposure to chronic stress disrupts blood-brain barrier integrity; still, little is known about intestinal barrier function in these conditions, particularly for the small intestine, where absorption of most foods and drugs takes place. Methods: We investigated how chronic social or variable stress, two mouse models of depression, impact the jejunum intestinal barrier in males and females. Mice were subjected to stress paradigms followed by analysis of gene expression profiles of intestinal barrier-related targets, fecal microbial composition, and blood-based markers. Results: Altered microbial populations and changes in gene expression of jejunum tight junctions were observed depending on the type and duration of stress, with sex-specific effects. We used machine learning to characterize in detail morphological tight junction properties, identifying a cluster of ruffled junctions in stressed animals. Junctional ruffling is associated with inflammation, so we evaluated whether lipopolysaccharide injection recapitulates stress-induced changes in the jejunum and observed profound sex differences. Finally, lipopolysaccharide-binding protein, a marker of gut barrier leakiness, was associated with stress vulnerability in mice, and translational value was confirmed on blood samples from women with MDD. Conclusions: Our results provide evidence that chronic stress disrupts intestinal barrier homeostasis in conjunction with the manifestation of depressive-like behaviors in a sex-specific manner in mice and, possibly, in human depression.

Characterization of protein aggregates in cream and skimmed human milk after heat and high-pressure pasteurization treatments.

Preservation processes applied to ensure microbial safety of human milk (HM) can modify the native structure of proteins and their bioactivities. Consequently, this study evaluated the effect of pasteurization methods (Holder pasteurization, high-temperature short-time (HTST), and high hydrostatic pressure (HHP)) of whole human milk (HM) on protein aggregates in skim milk and cream fractions. For heat-treated whole milk, insoluble protein aggregates at milk fat globule membrane (MFGM) were formed by disulfide and non-covalent bonds, but insoluble skim milk protein aggregates were only stabilized by non-covalent interactions. Contrary to heat treatment, the insolubilization of main proteins at the MFGM of HHP-treated HM was only through non-covalent interactions rather than disulfide bonds. Moreover, only heat treatment induced the insoluble aggregation of ⍺-lactalbumin. Overall, compared to heat treatment, HHP produced a milder effect on protein aggregation, validating the use of this process to better preserve the native state of HM bioactive proteins.

Pulse Globulins 11S and 7S: Origins, Purification Methods, and Techno-functional Properties.

A growing interest in pulse proteins in recent years results from their crucial role in the transition toward sustainable food systems. Consequently, current research is mainly focused on the production of protein ingredients and the evaluation of their nutritional and techno-functional properties for the development of animal product analogues. However, the individual impacts of the major proteins 11S legumin and 7S vicilin on pulse techno-functionalities remains unclear. Thus, this review aims to represent current knowledge on pulse 11S and 7S globulin origins, extraction, separation, and purification methods as well as their techno-functionalities. This paper also discusses the principal challenges related to pulse vicilin and legumin purification methods, such as efficiency and environmental concerns, as well as 11S/7S ratio variability. This review highlights the fact that 11S and 7S fractions serve different purposes in pulse functionality and that more efficient and eco-friendly purification techniques are required to properly assess their respective functional attributes. Such research would allow the determination of optimal 11S/7S ratios for the integration of pulse protein ingredients in various food formulations. Hence, food industries would be able to select species/varieties, agronomical methods, and processing methods to produce ingredients with suitable 11S/7S ratios, catering to consumers' ethical, environmental, and nutritional concerns.

Development of a human milk protein concentrate from donor milk: Impact of the pasteurization method on static in vitro digestion in a preterm newborn model.

The impact of high temperature short time (HTST, 72 °C, 15 s), Holder pasteurization- (63 °C, 30 min) and high hydrostatic pressure (HHP, 600 MPa-10 min) was evaluated on the digestibility of human milk protein concentrate (HMPC) by using a static in vitro gastrointestinal digestion system. The results showed that the processing steps used to produce the HMPC induced a decrease in readily available nitrogen (non-protein nitrogen and peptides). Overall, digestibility was similar between pasteurized and raw HMPC (degree of hydrolysis ranged from 26 to 34 %). Lactoferrin was more susceptible to gastric and intestinal digestion after thermal pasteurization. Additionally, the resistance of ß-casein to digestion increased after HHP and Holder pasteurization due to aggregation and changes in protein structure. During intestinal digestion, Holder pasteurization induced a higher release of arginine, phenylalanine and tyrosine from HMPC compared to raw and HHP-treated HMPC. Overall, protein structural changes induced by human milk (HM) processing (freeze-thawing and filtration) and pasteurization treatments affected HMPC proteolysis during in vitro digestion. However, protein digestion behaviors were quite similar for raw and HHP-treated HMPC compared to the thermal-treated HMPC, with no effect on lactoferrin digestion. Consequently, pasteurization of HMPC by HHP represents an interesting non-thermal process that preserves the HM bioactive proteins during digestion.

Donor human milk processing and its impact on infant digestion: A systematic scoping review of in vitro and in vivo studies.

When there is an inadequate supply of mother's milk, pasteurized donor human milk is preferred over formula to supplement feeds for preterm infants. Although providing donor milk helps to improve feeding tolerance and reduce necrotizing enterocolitis, changes to its composition and reductions in bioactivity during processing, are thought to contribute to the slower growth often exhibited by these infants. To improve the clinical outcomes of recipient infants by maximizing the quality of donor milk, research is currently investigating strategies to optimize all aspects of processing, including pooling, pasteurization, and freezing; however, reviews of this literature typically only summarize the impact of a processing technique on composition or bioactivity. Reviews of published research investigating the impact of donor milk processing on infant digestion/absorption are lacking and thus, was the objective for this systematic scoping review, Open Science Framework (https://doi.org/10.17605/OSF.IO/PJTMW). Databases were searched for primary research studies evaluating donor milk processing for pathogen inactivation or other rationale and subsequent effect on infant digestion/absorption. Non-human milk studies or those assessing other outcomes were excluded. Overall, 24 articles from 12,985 records screened were included. Most studied thermal methods to inactivate pathogens, predominantly Holder pasteurization (HoP) (62.5°C, 30 min) and high-temperature short-time. Heating consistently decreased lipolysis and increased proteolysis of lactoferrin and caseins; however, protein hydrolysis was unaffected from in vitro studies. The abundance and diversity of released peptides remain unclear and should be further explored. Greater investigation into less-harsh methods for pasteurization, such as high-pressure processing, is warranted. Only 1 study assessed the impact of this technique and found minimal impact on digestion outcomes compared with HoP. Fat homogenization appeared to positively impact fat digestion (n = 3 studies), and only 1 eligible study investigated freeze-thawing. Identified knowledge gaps regarding optimal methods of processing should be further explored to improve the quality and nutrition of donor milk.

Digestion of human milk processed by high pressure processing and Holder pasteurization using a dynamic in vitro model of the preterm infant.

Holder pasteurization (HoP) (62.5 °C, 30 min) of donor human milk is widely used to inactivate potential pathogens but may lead to denaturation and aggregation of bioactive proteins, reducing their functionality. In contrast, high pressure processing (HPP) is a non-thermal technique that minimally affects assessed bioactive components; however, it is unclear how HPP affects protein digestion, and retention of functional bioactive proteins. Raw or processed (HoP; HPP[500 MPa,10 min]) pools of milk (N = 3, from 9 donors) were subjected in triplicate to in vitro digestion simulating the preterm infant gastrointestinal tract. Compared to raw or HPP, HoP increased intestinal proteolysis of lactoferrin and bioactive milk fat globule membrane proteins. Lysozyme activity was impacted by digestion following HoP (72 % to 7 %)-significantly more than HPP (75 % to 34 %) or raw (100 % to 39 %), which did not differ. Proteins in HPP-treated donor milk are digested no different than raw milk, while preserved bioactivity remains functional upon digestion.

Gestational Diabetes Mellitus, Human Milk Composition, and Infant Growth.

Background: Gestational diabetes mellitus (GDM) is known to affect human milk composition. Aims of this study were to compare macronutrient and energy content of human milk of women with (GDM+) and without GDM (GDM-), to assess the association between maternal health and human milk macronutrient and energy content and association between human milk macronutrient and energy content and infant growth. Study Design and Methods: Two months after delivery, hindmilk samples were collected. Triglyceride (TG), lactose, and protein content of human milk were measured. An oral glucose tolerance test was performed. Infant weight and length at birth and 2 months were collected. Weight-for-age (WAZ) and weight-for-length z-scores were calculated. Results: Twenty-four GDM+ and 29 GDM- women were included. Protein, lactose, and energy content of human milk were similar between groups. TG concentration was higher in GDM+ than in GDM- women (6.3 ± 2.0 versus 5.3 ± 1.2, p = 0.04). This difference was no longer significant after adjustment for maternal age and infant sex (p = 0.23). Maternal age was associated with TG (r = 0.28, p = 0.04) and lactose (r = -0.30, p = 0.03), while fasting glucose was associated with proteins (r = 0.30, p = 0.03) and tended to be associated with TG (r = 0.27, p = 0.05) and energy (r = 0.24, p = 0.08). TG levels in human milk were associated with weight (ß: 0.26, 95% confidence interval [CI]: 0.02 to 0.50) and WAZ (ß: 0.40, 95% CI: 0.05 to 0.75) at 2 months among children unexposed (GDM-) to GDM, but not among children exposed (GDM+) Conclusions: In conclusion, GDM status, maternal age, and fasting glucose level were associated with human milk composition. Finally, TG in human milk was associated with infant growth among GDM- children but not among GDM+ children. ClinicalTrials.gov NCT02872402.

Effects of hexane on protein profile and techno-functional properties of pea protein isolates.

The defatting of legume flours with hexane is usually the first step in producing protein-rich ingredients. However, its impact on protein profiles, zeta potential, surface hydrophobicity and techno-functionality of pea proteins has not been evaluated. Consequently, this work aimed to evaluate the impact of the hexane defatting step on pea protein profiles, surface hydrophobicity and zeta potential, as well as techno-functional properties of non-defatted and defatted pea protein isolates. The results showed that alkaline extraction of hexane-defatted pea flour increased the net surface charge (zeta-potential) and reduced particle size of the pea protein isolate. Moreover, only the foaming properties of pea protein isolate generated from defatted pea flour were improved. Consequently, except for improving foaming properties, the defatting step is not essential for the production of pea protein isolate.

Candida krusei is the major contaminant of ultrafiltration and reverse osmosis membranes used for cranberry juice production.

Ultrafiltration (UF) and reverse osmosis (RO) are commonly used for the clarification and concentration of fruit juices. However, one of the main limitations of filtration membranes is biofouling, which reduces membrane efficiency and can contaminate the filtered product and lead to spoilage. In this study, the microbial fouling layers of UF and RO membranes from a Canadian cranberry juice processing plant were characterized. Unlike the microbiota found in cranberry juice, which is dominated by Bacillus sp. and other bacteria, both UF and RO membranes were mainly colonized by several strains of the yeast Candida krusei. A variation in bacterial and yeasts count was observed between tubular UF and spiral-wound RO membranes, and the analysis of the spatial distribution highlighted the homogeneity of the contamination across each membrane. Surprisingly, RO membranes had a higher level of contamination when compared to UF membranes. Furthermore, six strains of C. krusei were further characterized through multilocus sequence typing analysis, five of which exhibited unique allelic profiles and two of which were found to contain a new TRP1 allele.

Impact of emulsifiers for the nanoencapsulation with maltodextrin of cannabis oil by spray drying on the physicochemical properties and bioaccessibility of cannabinoids.

Our study aimed to investigate the impact of various emulsifiers, namely whey protein isolate (WPI), soy protein isolate (SPI), and Tween 80 (Tw), on their ability to encapsulate cannabis oil with maltodextrin as the wall material. The physicochemical properties of the powder, the stability of the cannabinoids, and their bioaccessibility during static in vitro digestion were examined. The average diameter of fat globules in liquid nanoemulsions was 170, 259, and 95 nm for WPI, SPI, and Tw, respectively. The encapsulation efficiency was high for protein emulsifiers (>95%) compared to Tw (∼16%). Upon powder reconstitution in water, the emulsified fat droplets remained stable for WPI (176 nm); however, higher fat globule size (diameters of 346 nm and 210 nm) was observed for SPI and Tw powders, respectively. All oil powders had high solubility (>97%). The peroxide value (PV) showed nearly a fourfold increase for the oil extracted from the powder than the initial PV of bulk oil (5.2 mEq). However, UPLC-TUV analysis of the main cannabinoids (CBD, THC, and CBN) indicated that there is no significant difference between the various formulations and the bulk oil, except for lower Tw. The in vitro digestion model results showed higher bioaccessibility of the cannabinoids for Tw (∼53%) than for proteins (WPI ∼ 7% and SPI ∼ 10%). These findings suggest that the emulsifiers used for spray drying nanoencapsulation of cannabis oil have an impact on the encapsulation efficiency and cannabinoid bioaccessibility, highlighting the importance of choosing adequate emulsifiying agents for their optimal oral delivery.

Factors affecting the modification of bovine milk proteins in high hydrostatic pressure processing: An updated review.

High hydrostatic pressure (HHP) treatment induces structural changes in bovine milk proteins depending on factors such as the temperature, pH, concentration, decompression rate, cycling, composition of the medium and pressure level and duration. An in-depth understanding of the impact of these factors is important for controlling HHP-induced modification of milk proteins and the interactions within or between them, which can be applied to prevent undesired aggregation, gelation, and precipitation during HHP processing or to obtain specific milk protein modifications to attain specific protein properties. In this regard, understanding the influences of these factors can provide insight into the modulation and optimization of HHP conditions to attain specific milk protein structures. In recent years, there has been a great research attention on HHP-induced changes in milk proteins influenced by factors such as pH, temperature, concentration, cycling, decompression condition, and medium composition. Hence, to provide insight into how these factors control milk protein structures under HHP treatment and to understand if their effects depend on HHP parameters and environmental conditions, this review discusses recent findings on how various factors (pH, temperature, cycling, decompression rate, medium composition, and concentration) affect HHP-induced bovine milk protein modification. Practical Application: The information provided in this review will be very useful to anticipate the challenges related to the formulation and development of pressure-treated milk and dairy products.

Preservation of Anti-cytomegalovirus Activity in Human Milk Following High-Pressure Processing Compared to Holder Pasteurization.

Pasteurized donor human milk is recommended for hospitalized preterm infants when mother's own milk is unavailable. Our aim was to compare the antiviral activity of human milk processed by Holder pasteurization (HoP) or high-pressure processing (HPP) against representative enveloped and non-enveloped viruses including cytomegalovirus and hepatitis A virus. Expressed milk from 20 donors collected from the Ontario Milk Bank was combined into 10 pools, each from two unique donors. Each pool was processed by HoP (62.5°C, 30 min) or HPP (500 MPa, 8 min, 4°C) and subsequently inoculated with cytomegalovirus or hepatitis A virus to achieve a final concentration of 5-log plaque-forming units/mL. Plaque reduction assays were used to quantify detectable virus after 30 min incubation (room temperature). Post hoc experiments using a 4 h incubation time were conducted if reductions were detected at 30 min. Irrespective of processing, cytomegalovirus concentrations declined in all pools after 30 min incubation (P < 0.0001). Milk processed by HoP exhibited significantly less reduction compared to raw milk (P = 0.0069). In post hoc experiments, anti-cytomegalovirus activity was maintained at 4 h, with high inter-pool variability. Hepatitis A virus concentration remained unchanged after 30 min incubation in raw and processed milk. Anti-cytomegalovirus activity in human milk is preserved following HoP and HPP, persisting up to 4 h post-inoculation; anti-hepatitis A virus activity was not observed in raw or processed milk. Further research is needed to understand how HoP or promising alternative processing methods affect the antiviral activity of donated milk, given its potential importance to recipient infants.

Effect of High Hydrostatic Pressure Intensity on Structural Modifications in Mealworm (Tenebrio molitor) Proteins.

Processing edible insects into protein extracts may improve consumer acceptability. However, a better understanding of the effects of food processing on the proteins is needed to facilitate their incorporation into food matrices. In this study, soluble proteins from Tenebrio molitor (10% w/v) were pressurized using high hydrostatic pressure (HHP) at 70-600 MPa for 5 min and compared to a non-pressurized control (0.1 MPa). Protein structural modifications were evaluated using turbidity measurement, particle-size distribution, intrinsic fluorescence, surface hydrophobicity, gel electrophoresis coupled with mass spectrometry, and transmission electron microscopy (TEM). The observed decrease in fluorescence intensity, shift in the maximum emission wavelength, and increase in surface hydrophobicity reflected the unfolding of mealworm proteins. The formation of large protein aggregates consisting mainly of hexamerin 2 and ⍺-amylase were confirmed by protein profiles on gel electrophoresis, dynamic light scattering, and TEM analysis. The typical aggregate shape and network observed by TEM after pressurization indicated the potential involvement of myosin and actin in aggregate formation, and these were detected by mass spectrometry. For the first time, the identification of mealworm proteins involved in protein aggregation phenomena under HHP was documented. This work is the first step in understanding the mealworm protein-protein interactions necessary for the development of innovative insect-based ingredients in food formulations.

Impact of Ultra-High Pressure Homogenization on the Structural Properties of Egg Yolk Granule.

Ultra-high pressure homogenization (UHPH) is a promising method for destabilizing and potentially improving the techno-functionality of the egg yolk granule. This study's objectives were to determine the impact of pressure level (50, 175 and 300 MPa) and number of passes (1 and 4) on the physico-chemical and structural properties of egg yolk granule and its subsequent fractions. UHPH induced restructuration of the granule through the formation of a large protein network, without impacting the proximate composition and protein profile in a single pass of up to 300 MPa. In addition, UHPH reduced the particle size distribution up to 175 MPa, to eventually form larger particles through enhanced protein-protein interactions at 300 MPa. Phosvitin, apovitellenin and apolipoprotein-B were specifically involved in these interactions. Overall, egg yolk granule remains highly stable during UHPH treatment. However, more investigations are needed to characterize the resulting protein network and to evaluate the techno-functional properties of UHPH-treated granule.

Health Benefits of Apple Juice Consumption: A Review of Interventional Trials on Humans.

Although numerous studies have reported the benefits of apple consumption on cardiometabolic health parameters and chronic disease prevention, few have focused on the effects of apple juice specifically. Juice consumption may be a convenient way to take advantage of the health effects of the bioactive components present in apples. The present review aims to summarize the current literature on health benefits of apple juice as reported in clinical trials in humans. Of the 67 studies retained, 20 interventional studies on humans were reviewed. Overall, cloudy apple juice consumption was found to be associated with several markers of cardiovascular health that may ultimately be relevant for cancer and neurodegenerative diseases. Most of the documentation was found regarding oxidative stress, as well as observations with other parameters such as markers of inflammation, lipid profile, and diabetes. This review suggests that, in 20 studies, apple juice consumed in moderation exerts positive effects on markers of cardiovascular disease risk (particularly on oxidative stress).

Impact of holder, high temperature short time and high hydrostatic pressure pasteurization methods on protein structure and aggregation in a human milk protein concentrate.

This work evaluated the impact of high temperature short time (HTST, 72 °C, 15 s), high hydrostatic pressure (HHP, 400-600 MPa at 5 and 10 min) and Holder pasteurization (HoP, 62.5 °C, 30 min) on protein profile and aggregation in a human milk protein concentrate (HMPC). The structural changes induced in milk proteins were investigated in HMPC as well as in sedimentable and non-sedimentable fractions recovered after ultracentrifugation. The results showed that heat treatments induced more protein denaturation and aggregation than did HHP treatments. Indeed, heat-induced protein aggregates observed in HMPC and the sedimentable fraction were mainly composed of lactoferrin and α-lactalbumin. More specifically, the concentration of lactoferrin in HMPC decreased by 86% after HTST and HoP whereas no effect was observed after HHP treatment. These results show the potential of HHP processing as a pasteurization method for HMPC since it minimizes the impact on protein structure, which generally correlates to protein quality and bioactivity.