Phosphoproteomics Revealed Differentially Expressed Sites and Function of the Bovine Milk Fat Globule Membrane in Colostrum and Mature Milk.

One type of large and intricate post-translational modification of milk proteins that has significant biological implications is phosphorylation. The characterization of phosphoproteins found in the bovine milk fat globule membrane (MFGM) is still mostly unknown. Here, label-free phosphoproteomics was used to identify 94 phosphorylation sites from 54 MFGM phosphoproteins in bovine colostrum (BC) and 136 phosphorylation sites from 91 MFGM phosphoproteins in bovine mature milk (BM). αs1-Casein and ß-casein were the most phosphorylated proteins in bovine colostrum. In bovine mature milk, perilipin-2 was the protein with the greatest number of phosphorylation sites. The results show that bovine colostrum MFGM phosphoproteins were mainly involved in immune function, whereas bovine mature MFGM phosphoproteins were mainly involved in metabolic function. Plasminogen and osteopontin were the most strongly interacting proteins in colostrum, whereas perilipin-2 was the most strongly interacting protein in bovine mature milk. This work demonstrates the unique alterations in the phosphorylation manner of the bovine MFGM protein during lactation and further expands our knowledge of the site characteristics of bovine MFGM phosphoproteins. This result confirms the value of MFGM as a reference ingredient for infant formula during different stages.

Human milk-derived versus bovine milk-derived fortifier use in very low birth weight infants: growth and vitamin D status.

Background: Human milk-derived fortifier (HMDF) coupled with human milk feeding in extremely premature infants reduces the adverse outcome risks of early exposure to bovine milk ingredients but may not provide enough nutrients for adequate catch-up growth compared with bovine milk-derived fortifier (BMDF). Objective: This study aims to compare HMDF and BMDF effects on growth parameters and serum 25-hydroxy vitamin D (25OHD) levels in preterm very low birth weight (VLBW) infants during the first 8 weeks of life. Methods: We present a retrospective chart review of inpatient VLBW infants with birth weight <1,500 g and gestational age <32 completed weeks who received either their mother's milk or donor breast human milk fortified with HMDF or BMDF for the first 8 weeks. Weight, head circumference, length gain, and 25OHD level were calculated at 4 and 8 weeks of age. Results: A total of 139 VLBW infants (91 HMDF + 48 BMDF) received fortified human milk without any supplemental premature formula from birth to 4 weeks of age, of whom 44 (37 HMDF + 7 BMDF) continued until 8 weeks of age. There was no statistically significant difference in the growth parameters between the two groups at 4 and 8 weeks of age. Serum 25OHD level in the HMDF group was significantly higher compared with that in the BMDF group at 4 weeks of age despite receiving lower vitamin D supplementation. Conclusion: Similar gain in growth parameters in HMDF and BMDF groups at 4 and 8 weeks of age was observed, suggesting that HMDF provides adequate nutrients for growth in VLBW infants. A higher 25OHD level in HMDF may suggest better absorption.

Application of Milk Exosomes for Musculoskeletal Health: Talking Points in Recent Outcomes.

Milk is a nutrient-rich food source, and among the various milks, breast milk is a nutrient source provided by mothers to newborns in many mammals. Exosomes are nano-sized membranous extracellular vesicles that play important roles in cell-to-cell communication. Exosomes originate from endogenous synthesis and dietary sources such as milk. Discovered through electron microscopy as floating vesicles, the existence of exosomes in human milk was confirmed owing to a density between 1.10 and 1.18 g/mL in a sucrose gradient corresponding to the known density of exosomes and detection of MHC classes I and II, CD63, CD81, and CD86 on the vesicles. To date, milk exosomes have been used for treating many diseases, including cancers, and are widely proposed as promising carriers for the delivery of chemotherapeutic agents. However, few studies on milk exosomes focus on geriatric health, especially sarcopenia and osteoporosis related to bone and muscle. Therefore, the present study focused on milk exosomes and their cargoes, which are potential candidates for dietary supplements, and when combined with drugs, they can be effective in treating musculoskeletal diseases. In this review, we introduce the basic concepts, including the definition, various sources, and cargoes of milk exosomes, and exosome isolation and characterization methods. Additionally, we review recent literature on the musculoskeletal system and milk exosomes. Since inflammation and oxidative stress underly musculoskeletal disorders, studies reporting the antioxidant and anti-inflammatory properties of milk exosomes are also summarized. Finally, the therapeutic potential of milk exosomes in targeting muscle and bone health is proposed.

Comparison of Whey Proteome and Glycoproteome in Bovine Colostrum and Mature Milk.

Milk samples were collected from 10 cows, in the colostrum (3-4 days) and mature (90 days) lactation stage, to assess the differential expression of all whey proteins and N-glycoproteins. In total, 240 whey proteins and 315 N-glycosylation sites on 214 glycoproteins were quantified. GO annotations, KEGG pathway analysis, and protein classification were performed to understand the similarities and differences of the biological functions of whey proteins and N-glycoproteins among different lactation stages in bovine milk. Furthermore, differential expression of whey proteins and whey N-glycosylated proteins was found between different lactation stages. The related changes of biological functions in differentially expressed proteins were discussed. For example, the increased frequency of glycosylation on lactoferrin and folate receptor alpha occurring in bovine colostrum may provide protection and stimulate development of the newborn calf. Our study thereby improves understanding of variations of glycosylation sites on milk glycoproteins among lactation stages.

Effect of dietary seaweed (Ascophyllum nodosum) supplementation on milk mineral concentrations, transfer efficiency, and hematological parameters in lactating Holstein cows.

This study investigated the effect of feeding seaweed (Ascophyllum nodosum) to dairy cows on milk mineral concentrations, feed-to-milk mineral transfer efficiencies, and hematological parameters. Lactating Holstein cows (n = 46) were allocated to 1 of 2 diets (n = 23 each): (1) control (CON; without seaweed) and (2) seaweed (SWD; replacing 330 g/d of dried corn meal in CON with 330 g/d dried A. nodosum). All cows were fed the CON diet for 4 wk before the experiment (adaptation period), and animals were then fed the experimental diets for 9 wk. Samples included sequential 3-wk composite feed samples, a composite milk sample on the last day of each week, and a blood sample at the end of the study. Data were statistically analyzed using a linear mixed effects model with diet, week, and their interaction as fixed factors; cow (nested within diet) as a random factor; and data collected on the last day of the adaptation period as covariates. Feeding SWD increased milk concentrations of Mg (+6.6 mg/kg), P (+56 mg/kg), and I (+1,720 µg/kg). It also reduced transfer efficiency of Ca, Mg, P, K, Mn, and Zn, and increased transfer efficiency of Mo. Feeding SWD marginally reduced milk protein concentrations, whereas there was no effect of SWD feeding on cows' hematological parameters. Feeding A. nodosum increased milk I concentrations, which can be beneficial when feed I concentration is limited or in demographics or populations with increased risk of I deficiency (e.g., female adolescents, pregnant women, nursing mothers). However, care should also be taken when feeding SWD to dairy cows because, in the present study, milk I concentrations were particularly high and could result in I intakes that pose a health risk for children consuming milk.

Quantitative phosphoproteome analysis reveals differential whey phosphoproteins of bovine milk during lactation.

Whey proteins in bovine milk, as the most widely used nutritional components for infant formulae, have been paid more attention. However, the phosphorylation of proteins in bovine whey during lactation has not been thoroughly researched. In this study, a total of 185 phosphorylation sites on 72 phosphoproteins were identified in bovine whey during lactation. 45 differentially expressed whey phosphoproteins (DEWPPs) in colostrum and mature milk were focused on by bioinformatics approaches. Gene Ontology annotation indicated that blood coagulation, extractive space, and protein binding played a key role in bovine milk. The critical pathway of DEWPPs was related to the immune system according to KEGG analysis. Our study investigated the biological functions of whey proteins from a phosphorylation perspective for the first time. The results elucidate and increase our knowledge of differentially phosphorylation sites and phosphoproteins in bovine whey during lactation. Additionally, the data might offer fresh insight into the development of whey protein nutrition.

Key changes in bovine milk immunoglobulin G during lactation: NeuAc sialylation is a hallmark of colostrum immunoglobulin G N-glycosylation.

We monitored longitudinal changes in bovine milk IgG in samples from four cows at 9 time points in between 0.5 and 28 days following calving. We used peptide-centric LC-MS/MS on proteolytic digests of whole bovine milk, resulting in the combined identification of 212 individual bovine milk protein sequences, with IgG making up >50 percent of the protein content of every 0.5 d colostrum sample, which reduced to ≤3 percent in mature milk. In parallel, we analyzed IgG captured from the bovine milk samples to characterize its N-glycosylation, using dedicated methods for bottom-up glycoproteomics employing product ion-triggered hybrid fragmentation; data are available via ProteomeXchange with identifier PXD037755. The bovine milk IgG N-glycosylation profile was revealed to be very heterogeneous, consisting of >40 glycoforms. Furthermore, these N-glycosylation profiles changed substantially over the period of lactation, but consistently across the four individual cows. We identified NeuAc sialylation as the key abundant characteristic of bovine colostrum IgG, significantly decreasing in the first days of lactation, and barely detectable in mature bovine milk IgG. We also report, for the first time to our knowledge, the identification of subtype IgG3 in bovine milk, alongside the better-documented IgG1 and IgG2. The detailed molecular characteristics we describe of the bovine milk IgG, and their dynamic changes during lactation, are important not only for the fundamental understanding of the calf's immune development, but also for understanding bovine milk and its bioactive components in the context of human nutrition.

The Texture of Camel Milk Cheese: Effects of Milk Composition, Coagulants, and Processing Conditions.

Numerous people in African, Middle Asian, Middle Eastern, and Gulf Cooperation Council (GCC) countries highly value camel milk (CM) as it plays a vital role in their diet. The protein composition of CM as well as the structure of its casein micelles differs significantly from bovine milk (BM). Cheeses made from CM have a weak curd and soft texture compared to those made from BM. This review article presents and discusses the effect of milk protein composition, processing conditions (pasteurization and high-pressure treatment), and coagulants (camel chymosin, organic acids, plant proteases) on the quality of CM cheeses. CM cheese's weak texture is due to compositional characteristics of the milk, including low κ-casein-to-ß-casein ratio (≈0.05 in CM vs. ≈0.33 in BM), large micelle size, different whey protein components, and higher proteolytic activity than BM. CM cheese texture can be improved by preheating the milk at low temperatures or by high pressure. Supplementing CM with calcium has shown inconsistent results on cheese texture, which may be due to interactions with other processing conditions. Despite their structure, CM cheeses are generally well liked in sensory studies.

Similarity Index for the Fat Fraction between Breast Milk and Infant Formulas.

The similarity of the fat fraction in infant formulas rich in either bovine milk fat (MF) or vegetable oil (VO) to breast milk was evaluated by analyzing their lipid composition. Milk fat-rich formulas were highly similar (average similarity index 0.68) to breast milk compared to the VO-rich formulas (average similarity index 0.56). The highest difference in the indices was found in the contents of cholesterol (0.66 vs 0.28 in MF- and VO-rich formulas, respectively, on average) and polar lipids (0.84 vs 0.53), the positional distribution of fatty acids in the sn-2 position of triacylglycerols (0.53 vs 0.28), and fatty acid composition (0.72 vs 0.54). The VO-based formulas were superior in similarity in n - 6 PUFA. Thus, the addition of bovine MF fractions is an effective way to increase the similarity between the lipid composition of infant formulas and human milk.

Carotenoids in Milk and the Potential for Dairy Based Functional Foods.

Carotenoids are a family of over 1100 known natural pigments synthesized by plants, algae, fungi and bacteria. Dietary intake of carotenoids is necessary for mammals as they cannot be synthesized in the body. In cows, the nature of the diet consumed strongly influences the composition of milk produced and this includes carotenoid concentration and profile. Fresh forage is the richest source of carotenoids for cows. The main carotenoids identified in forages are lutein, ß-carotene, zeaxanthin and epilutein. Manipulating cow feed via carotenoid supplementation increases the carotenoid content of bovine milk. In humans, carotenoids have anti-oxidant, anti-inflammatory and provitamin A activity. Lutein is a major carotenoid in human milk and the brain tissue of adults and infants. Lutein and zeaxanthin are linked to improved eye health and cognitive function. Traditionally for humans, fruit and vegetables have been the main source of carotenoid intake. Functional foods present an opportunity to incorporate these naturally occurring compounds into milk products for added health benefits, widening the range of dietary sources of carotenoids. We offer an overview of the literature to date on carotenoid-fortified dairy products and infant formula. This review will describe and summarize the key mechanisms by which the carotenoid profile of bovine milk can be manipulated. We present findings on the origin and role of carotenoids in bovine and human milk, outline factors that impact the carotenoid content of milk, evaluate carotenoid-fortified milk products and discuss the associated challenges, such as bioaccessibility and stability.

Short communication: The effect of pectin and sodium alginate on labans made from camel milk and bovine milk.

Camel milk (CM) is gaining scientific attention due to its potential health and therapeutic benefits. Fermented drinkable yogurts (labans) were prepared from CM and bovine milk (BM) using mixed Streptococcus thermophilus and Lactobacillus delbrueckii ssp. bulgaricus bacteria supplemented with 1 of 2 hydrocolloids: pectin (0.1-0.3%) or sodium alginate (0.1-0.5%). The different labans were compared by studying their acidity and rheology as well as their structural and sensory properties. The CM and BM labans had titratable acidity values that ranged from 0.85 to 1.27 and 0.61 to 0.93%, respectively. Pectin at 0.2% enhanced the rheological properties of BM labans, but had no effect in CM labans. Sodium alginate at 0.3% and 0.5% increased viscosity, elastic or storage modulus (G'), and viscous or loss modulus (G″) values for both types of laban. Scanning electron microscopy indicated that the CM laban contained lower levels of "spike-like structures" than BM laban, and that the addition of hydrocolloids improved this effect. Quantitative descriptive sensory analysis showed that CM labans fortified with either 0.2% pectin or 0.3% sodium alginate were comparable to commercial BM laban in viscous mouthfeel. Fortified CM labans were more acidic and had stronger flavors than unfortified samples. Overall, this study demonstrated that the addition of sodium alginate or pectin at intermediate levels permits production of palatable CM labans of a satisfactory viscous consistency.

Comprehensive Assessment of Essential and Potentially Toxic Trace Elements in Bovine Milk and Their Feeds in Different Agro-climatic Zones of Sri Lanka.

Fresh milk is an important source of essential mineral supplement for humans. However, the levels of trace elements in milk are an important component of its safety and quality. Trace elements also act as a bio-indicator of agricultural pollution. Samples of raw cow milk (n = 68), animal feed [forage (n = 36) and concentrates (n = 14)], and water (n = 35) were collected from different agro-climatic regions of Sri Lanka. The concentrations of 15 trace elements including toxic heavy metals such as Cd, Pb, and As were quantified using inductively coupled plasma mass spectrometry after microwave-assisted digestion. Among the studied trace elements, the mean elemental concentrations of Se, Cd, As, and Cu in cow milk were 18.1, 1.45, 7.35, and 71.7 µg L-1, respectively. The mean concentrations of these elements in forage were 0.216, 0.066, 0.046, and 9.21 mg kg-1, and in concentrate feed were 0.329, 0.202, 0.229, and 2.28 mg kg-1, respectively. The As content of the studied milk was well below the maximum permissible level while 15% of the samples had Pb exceeding the permissible limit of 20 µg L-1. However, As, Cd, and Pb levels in drinking water provided to animals were well below the WHO permissible limits. The results of this study revealed that the trace elements in cow milk depend mainly on the feed, and hence, levels of these contaminants in feed materials should be monitored. Although due to the consumption of milk, the populations have a minor exposure to trace elements and also minimum health risks, a detailed study with a large number of samples is highly recommended.

Evaluation of bovine milk extracellular vesicles for the delivery of locked nucleic acid antisense oligonucleotides.

The natural capacity of extracellular vesicles (EVs) to transport their payload to recipient cells has raised big interest to repurpose EVs as delivery vehicles for xenobiotics. In the present study, bovine milk-derived EVs (BMEVs) were investigated for their potential to shuttle locked nucleic acid-modified antisense oligonucleotides (LNA ASOs) into the systemic circulation after oral administration. To this end, a broad array of analytical methods including proteomics and lipidomics were used to thoroughly characterize BMEVs. We found that additional purification by density gradients efficiently reduced levels of non-EV associated proteins. The potential of BMEVs to functionally transfer LNA ASOs was tested using advanced in vitro systems (i.e. hPSC-derived neurons and primary human cells). A slight increase in cellular LNA ASO internalization and target gene reduction was observed when LNA ASOs were delivered using BMEVs. When dosed orally in mice, only a small fraction (about 1% of total administered dose) of LNA ASOs was recovered in the peripheral tissues liver and kidney, however, no significant reduction in target gene expression (i.e. functional knockdown) was observed.

Effects of Industrial Thermal Treatments on the Release of Bovine Colostrum Glycoprotein N-Glycans by Endo-ß-N-acetylglucosaminidase.

N-Glycans are structurally similar to human milk oligosaccharides, the gold standard prebiotics for infants. Bovine milk N-glycans released by endo-ß-N-acetylglucosaminidase (EndoBI-1) were shown to have similar prebiotic selectivity as human milk oligosaccharides, explaining the interest for N-glycan recovery for use as prebiotics. Industrial thermal treatments such as high-temperature short-time (HTST) and ultra-high-temperature (UHT) might favor the enzymatic deglycosylation of N-glycans through promoting protein denaturation. We investigated the effects of HTST (72 °C for 15 s) and UHT (135 °C for 3 s) on N-glycan release from bovine colostrum glycoproteins by nonimmobilized and amino-immobilized EndoBI-1. A total of 104 N-glycans including isomers/anomers were identified by high-resolution mass spectrometry. In both EndoBI-1 forms, HTST increased the release of N-glycans; however, the impact of UHT on releasing N-glycans was comparable to the nonthermal treatment. Although the amino-immobilized enzyme similarly released neutral N-glycans as the free form, it released fewer sialylated and fucosylated N-glycans.

Characterization and comparison of lipids in bovine colostrum and mature milk based on UHPLC-QTOF-MS lipidomics.

Lipids in bovine milk have several biological activities, with implications for human health and the physical functionality of foods. However, alterations in the lipid profile of bovine milk during lactation are not well-studied. This study aimed to identify differences in lipids between bovine colostrum and mature milk, using a lipidomics approach. Using an advanced mass spectrometry-based quantitative lipidomics approach, 335 lipids assigned to 13 subclasses were characterized in bovine colostrum (BC) and mature milk (BM). In total, 63 significantly differential lipids (SDLs) were identified. Among the 63 SDLs, the levels of 21 lipids were significantly lower in BM than in BC, including 5 glycerophosphatidylethanolamines (PEs), 1 glycerophosphatidylglycerol (PG), and 15 triacylglycerols (TGs). The levels of the remaining 42 lipids increased in BM, including 1 cardiolipin (CL), 9 diacylglycerols (DGs), 9 dihexosylceramides (Hex2Cers), 3 hexosylceramides (HexCers), 3 glycerophosphatidic acids (PAs), 2 glycerophosphatidylcholines (PCs), 12 PEs, and 3 TGs. Furthermore, the correlations and related metabolic pathways of these 63 SDLs were analyzed to explore the mechanisms that alter bovine milk lipids during lactation. The seven most relevant pathways identified herein, ranked in accordance with their degree of influence on lactation, were glycerophospholipid metabolism, sphingolipid metabolism, glycerolipid metabolism, glycosylphosphatidylinositol-anchor biosynthesis, linoleic acid metabolism, alpha-linolenic acid metabolism, and arachidonic acid metabolism. Our results provide essential insights into mechanisms underlying alterations in bovine milk lipids during different lactation periods, along with practical information of specific nutrition and quality assessments for the dairy industry.

Lactoferrin from Bovine Milk: A Protective Companion for Life.

Lactoferrin (Lf), an iron-binding multifunctional glycoprotein belonging to the transferrin family, is present in most biological secretions and reaches particularly high concentrations in colostrum and breast milk. A key function of lactoferrin is non-immune defence and it is considered to be a mediator linking innate and adaptive immune responses. Lf from bovine milk (bLf), the main Lf used in human medicine because of its easy availability, has been designated by the United States Food and Drug Administration as a food additive that is generally recognized as safe (GRAS). Among the numerous protective activities exercised by this nutraceutical protein, the most important ones demonstrated after its oral administration are: Antianemic, anti-inflammatory, antimicrobial, immunomodulatory, antioxidant and anticancer activities. All these activities underline the significance in host defence of bLf, which represents an ideal nutraceutical product both for its economic production and for its tolerance after ingestion. The purpose of this review is to summarize the most important beneficial activities demonstrated following the oral administration of bLf, trying to identify potential perspectives on its prophylactic and therapeutic applications in the future.

Urinary Metabolomic Profile of Preterm Infants Receiving Human Milk with Either Bovine or Donkey Milk-Based Fortifiers.

Fortification of human milk (HM) for preterm and very low-birth weight (VLBW) infants is a standard practice in most neonatal intensive care units. The optimal fortification strategy and the most suitable protein source for achieving better tolerance and growth rates for fortified infants are still being investigated. In a previous clinical trial, preterm and VLBW infants receiving supplementation of HM with experimental donkey milk-based fortifiers (D-HMF) showed decreased signs of feeding intolerance, including feeding interruptions, bilious gastric residuals and vomiting, with respect to infants receiving bovine milk-based fortifiers (B-HMF). In the present ancillary study, the urinary metabolome of infants fed B-HMF (n = 27) and D-HMF (n = 27) for 21 days was analyzed by 1H NMR spectroscopy at the beginning (T0) and at the end (T1) of the observation period. Results showed that most temporal changes in the metabolic responses were common in the two groups, providing indications of postnatal adaptation. The significantly higher excretion of galactose in D-HMF and of carnitine, choline, lysine and leucine in B-HMF at T1 were likely due to different formulations. In conclusion, isocaloric and isoproteic HM fortification may result in different metabolic patterns, as a consequence of the different quality of the nutrients provided by the fortifiers.

Comparison of the hypoglycemic and antithrombotic (anticoagulant) actions of whole bovine and camel milk in streptozotocin-induced diabetes mellitus in rats.

People with diabetes are at higher risk of fatal thromboembolic accidents in the cerebral and coronary circulations, especially stroke and ischemic heart disease. We have previously described the hypoglycemic, hypolipidemic, and anticoagulant activity of orally administered camel milk in streptozotocin-induced diabetic rats. In the present study in the same animal model, we extended these observations by comparing camel milk and the more available and widely consumed bovine milk with respect to their antidiabetic and antithrombotic actions. Rats were rendered diabetic by intraperitoneal streptozotocin (65 mg/kg), and then camel milk or bovine milk was administered orally for 8 wk. We evaluated the changes in body weight, fasting blood glucose, glucose tolerance, blood coagulation profile, and platelet function. Diabetic rats developed weight loss, hyperglycemia, glucose intolerance, inhibition of platelet aggregation responses to arachidonic acid and adenosine diphosphate, a marked decrease (>50%) in plasma fibrinogen levels, and short activated partial thromboplastin time. Treatment with camel milk or bovine milk reversed these abnormalities, resulting in weight gain, decreased blood glucose levels, and improved glucose tolerance. Despite the more remarkable antidiabetic action of camel milk, treatment with bovine milk was more effective in correcting plasma fibrinogen levels and restoring inhibited platelet aggregation responses. Long-term administration of camel milk or bovine milk counteracted streptozotocin-induced metabolic manifestations in rats, maintained platelet function, and abolished coagulopathy-associated fibrinogen consumption. Notably, the antidiabetic effect of camel milk was more pronounced than that of bovine milk, but bovine milk exhibited more potent anticoagulant activity than camel milk. These findings should encourage further clinical trials to assess the efficiency of camel milk and bovine milk or their derived peptides as food supplements or potential nonpharmacological therapies for dysglycemia and the vascular complications of diabetes mellitus.

FortiColos - a multicentre study using bovine colostrum as a fortifier to human milk in very preterm infants: study protocol for a randomised controlled pilot trial.

BACKGROUND: Very preterm infants (< 32 weeks gestation) have a relatively high nutrient requirement for growth and development. The composition of human milk is often inadequate to ensure optimal growth so it is common to fortify human milk for very preterm infants with nutrient fortifiers based on bovine milk. However, there are concerns that bovine milk-based fortifiers may increase the risk of feeding intolerance, necrotizing enterocolitis and late-onset sepsis. We hypothesize that a bovine colostrum-based product is a suitable alternative to bovine milk-based products when used as a fortifier to human milk in very preterm infants. METHODS/DESIGN: In an open-label multicentre randomised controlled pilot trial, 200 very preterm infants (26 + 0 to 30 + 6 weeks gestation at birth) will be randomly allocated to a bovine colostrum-based or a bovine milk-based fortifier added to mother's own milk and/or human donor milk. Outcomes are growth rate, incidence of necrotizing enterocolitis and late-onset sepsis, a series of paraclinical endpoints, and practical feasibility of using the novel fortifier for very preterm infants. DISCUSSION: The optimal enteral diet and feeding regimen for very preterm infants remain debated; this clinical trial will document the feasibility, safety and preliminary efficacy of using bovine colostrum, rich in nutrients and bioactive factors, as a novel fortifier for human milk to very preterm infants. Data on infant growth, metabolism, gut function and immunity will be assessed from clinical data as well as blood and stool samples. TRIAL REGISTRATION: Registered retrospectively 25 May 2018 at ClinicalTrials.gov: NCT03537365 .

Antiulcerative and Antinociceptive Activities of Casein and Whey Proteins.

The aim of this study was to evaluate the antiulcerative and antinociceptive activities of milk proteins using the induced gastric ulcer with ethanol rat model and the acetic acid-induced writhing mouse model. Casein (CN), (100, 300, and 1000 mg kg-1) doses presented antiulcerative activity on a dose-dependent manner with values of 30.8%, 41.4%, and 57.0% of inhibition measured using the ulcerative lesions index (ULI), respectively. Whey protein concentrate (WPC), (100, 300, and 1000 mg kg-1) doses presented antiulcerative activity on a dose-dependent manner with values of 48.9%, 65.5%, and 68.22% of ULI inhibition, respectively. CN, casein hydrolysates (CNH), WPC, and whey protein hydrolysates (WPH), (3, 10, and 30 mg kg-1) doses presented antinociceptive activity using the acetic acid-induced writhing in the mouse model. CN (30 mg kg-1) presented a value of 40% of inhibition writhing, and CNH (30 mg kg-1) presented antinociceptive activity with a value up to 46% of writhing inhibition. WPC (30 mg kg-1) presented a value of 52.50%, and WPH (30 mg kg-1) presented antinociceptive activity with a value up to 88.00% of writhing inhibition. In conclusion, CN and WPC demonstrated in vivo antiulcerative properties and represent a promising alternative to be used as protectors of the gastric mucosa. CNH and WPH demonstrated in vivo antiulcerative properties and represent a promising alternative to be used as natural analgesic.