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20 pages, 3939 KiB  
Article
Structural Characterization and Abundance of Sialylated Milk Oligosaccharides in Holstein Cows during Early Lactation
by Lisa Isernhagen, Christina E. Galuska, Andreas Vernunft and Sebastian P. Galuska
Foods 2024, 13(16), 2484; https://doi.org/10.3390/foods13162484 - 7 Aug 2024
Viewed by 307
Abstract
Among other bioactive molecules, milk contains high amounts of sialylated milk oligosaccharides (MOs) that influence numerous processes in the offspring. For instance, sialylated MOs inhibit the invasion of pathogens and positively influence the gut microbiome to support the optimal development of the offspring. [...] Read more.
Among other bioactive molecules, milk contains high amounts of sialylated milk oligosaccharides (MOs) that influence numerous processes in the offspring. For instance, sialylated MOs inhibit the invasion of pathogens and positively influence the gut microbiome to support the optimal development of the offspring. For these reasons, sialylated MOs are also used in infant formula as well as food supplements and are potential therapeutic substances for humans and animals. Because of the high interest in sialylated bovine MOs (bMOs), we used several analytical approaches, such as gas and liquid chromatography in combination with mass spectrometry, to investigate in detail the profile of sialylated bMOs in the milk of Holstein Friesian cows during early lactation. Most of the 40 MOs identified in this study were sialylated, and a rapid decrease in all detected sialylated bMOs took place during the first day of lactation. Remarkably, we observed a high variance within the sialylation level during the first two days after calving. Therefore, our results suggest that the content of sialylated MOs might be an additional quality marker for the bioactivity of colostrum and transitional milk to ensure its optimized application for the production of milk replacer and food supplements. Full article
Show Figures

Figure 1

Figure 1
<p>Quantification of Neu5Gc in milk from Holstein Friesian cows. (<b>A</b>) Sampling timeline for milk collection. The time ranges of colostrum, transitional milk, and mature milk were specified on the basis of Silva et al. [<a href="#B49-foods-13-02484" class="html-bibr">49</a>]. Created with BioRender.com. (<b>B</b>) Scheme of the DMB-RP-HPLC strategy for sialic acid quantification. Sialic acid residues in milk as well as in the extracted bMOs fraction were released by hydrolysis and subsequently labeled with DMB for fluorescence detection using an RP-HPLC system equipped with a fluorescence detector. Created with BioRender.com. (<b>C</b>) Box and whisker plots (median; min to max) showing the Neu5Gc values during early lactation (n = 6 animals) as well as the values for Neu5Ac (<b>D</b>) and Neu5Gc (<b>E</b>) in the extracted bMOs fraction (n = 5 animals). The statistical analysis and graphs were generated using BioRender.com. Significant differences are denoted as follows: *: <span class="html-italic">p</span> &lt; 0.05, **: <span class="html-italic">p</span> &lt; 0.01, ***: <span class="html-italic">p</span> &lt; 0.001, ****: <span class="html-italic">p</span> &lt; 0.0001.</p>
Full article ">Figure 2
<p>Analysis of bMO distribution using LC-MS. (<b>A</b>) Scheme of bMO analysis using LC-MS. After bMO enrichment using PGC cartridges, the resulting bMOs were analyzed using HILIC-HESI-MS/(MS). Created with BioRender.com. (<b>B</b>) The displayed MS fragments of bMOs were used for the identification of bMOs. For the manual verification process, at least one of those fragments had to be found in the corresponding MS<sup>2</sup> spectrum to identify the detected bMO. For the sialylated bMOs, the most significant fragment was always the cleaved sialic acid residues (Neu5Ac or Neu5Gc). The bMO structures were computed with GlycoWorkbench 2 [<a href="#B43-foods-13-02484" class="html-bibr">43</a>].</p>
Full article ">Figure 3
<p>Proposed structures of the detected disaccharides lactose and lactosamine in addition to bMOs in Holstein Friesian cows during early lactation. For some bMOs, multiple linkages and compositions have been reported. Due to the lack of specific linkage analysis and for clarity purposes, not all possible isomers are visualized. The bMO structures were designed with GlycoWorkbench 2 [<a href="#B43-foods-13-02484" class="html-bibr">43</a>] and assembled using BioRender.com. Each structure is given the corresponding number used in this study, which is also described in <a href="#foods-13-02484-t001" class="html-table">Table 1</a>.</p>
Full article ">Figure 4
<p>LC-MS analysis of sialyllactose (SL). (<b>A</b>) Extracted ion chromatogram (EIC) of SLs shown for representative samples on the day of calving. (<b>B</b>) MS spectrum at 5.46 min showing the deprotonated molecular ion [M-H]<sup>−</sup> of SL with its corresponding MS/MS spectrum (<b>D</b>) showing two MO fragments as a cross-ring fragment of a Hex with 87.0072 <span class="html-italic">m</span>/<span class="html-italic">z</span> and a detached Neu5Ac at 290.0884 <span class="html-italic">m</span>/<span class="html-italic">z</span>. (<b>C</b>) MS spectrum at 6.68 min showing the deprotonated molecular ion [M-H]<sup>−</sup> of SL with its corresponding MS/MS spectrum (<b>E</b>) showing fragments at 87.0072 and 290.0883 <span class="html-italic">m</span>/<span class="html-italic">z</span>, as well as Neu5Ac attached to a Hex (470.1522 <span class="html-italic">m</span>/<span class="html-italic">z</span>) and Neu5Ac attached to a Hex with an additional cross-ring fragment of another Hex (572.1846 <span class="html-italic">m</span>/<span class="html-italic">z</span>). The bMO structures were designed with GlycoWorkbench 2 [<a href="#B43-foods-13-02484" class="html-bibr">43</a>].</p>
Full article ">Figure 5
<p>LC-MS analysis of DSL. (<b>A</b>) MS spectrum at 10.99 min showing the deprotonated molecular ion for DSL containing a Neu5Ac dimer attached to the lactose core and the corresponding MS<sup>2</sup> spectrum for DSL at this time. The MS<sup>2</sup> spectrum shows three fragments, two of which are already mentioned for SL in <a href="#foods-13-02484-f004" class="html-fig">Figure 4</a> (87.0072 and 290.0881 <span class="html-italic">m</span>/<span class="html-italic">z</span>), and one larger fragment is SL with 632.2120 <span class="html-italic">m</span>/<span class="html-italic">z</span>, validating the linear composition of DSL. (<b>B</b>) MS spectrum at 11.51 min showing the deprotonated molecular ion for heterogenic DSL (hDSL) consisting of Neu5Ac as well as Neu5Gc attached to the lactose core and the corresponding MS<sup>2</sup> spectrum for hDSL at this time. The MS<sup>2</sup> spectrum shows four fragments, two of which were already mentioned for SL in <a href="#foods-13-02484-f004" class="html-fig">Figure 4</a> (87.0071 and 290.0885 <span class="html-italic">m</span>/<span class="html-italic">z</span>), as well as one larger fragment, NGL, with 648.2013 <span class="html-italic">m</span>/<span class="html-italic">z</span>, validating the linear composition of hDSL as NGL with an additional Neu5Ac attached. (<b>C</b>) The MS spectrum at 12.69 min displays the deprotonated molecular ion for hDSL and the corresponding MS<sup>2</sup> spectrum for hDSL at this time. The MS<sup>2</sup> spectrum shows four fragments, three of which were already mentioned for hDSL (87.0071, 290.0872, and 306.0837 <span class="html-italic">m</span>/<span class="html-italic">z</span>), as well as SL with 632.2031 <span class="html-italic">m</span>/<span class="html-italic">z</span>, validating the linear composition of the second hDSL as SL with an additional Neu5Gc attached. The bMO structures were designed with GlycoWorkbench 2 [<a href="#B43-foods-13-02484" class="html-bibr">43</a>].</p>
Full article ">Figure 6
<p>LC-MS analysis of sialyllactosamine (SLN). (<b>A</b>) Extracted ion chromatogram (EIC) of SLN shown for a representative sample on the day of calving. (<b>B</b>) MS spectrum at 5.10 min showing the deprotonated molecular ion 673.2306 <span class="html-italic">m</span>/<span class="html-italic">z</span> of SLN with the corresponding MS<sup>2</sup> spectrum, (<b>D</b>) which displays the same fragments as those displayed for SL in <a href="#foods-13-02484-f005" class="html-fig">Figure 5</a>, with an additional fragment of another cross-ring fragment of Hex at 170.0449 <span class="html-italic">m</span>/<span class="html-italic">z</span>. (<b>C</b>) MS spectrum at 5.50 min showing the deprotonated molecular ion 673.2311 <span class="html-italic">m</span>/<span class="html-italic">z</span> of SLN with the corresponding MS<sup>2</sup> spectrum, (<b>E</b>) which shows the same fragments as noted at 5.10 min. Additionally, there is another fragment of Neu5Ac attached to a cross-ring fragment of a Hex (306.1190 <span class="html-italic">m</span>/<span class="html-italic">z</span>), which indicates the α2,6-linkage of Neu5Ac. The bMO structures were computed with GlycoWorkbench 2 [<a href="#B43-foods-13-02484" class="html-bibr">43</a>].</p>
Full article ">Figure 7
<p>Distribution of bMOs during lactation. (<b>A</b>) The total ion chromatography (TIC) from the colostrum samples on day 0 p.p. of the analyzed Holstein cattle were overlaid using FreeStyle software (Thermo Fisher). The MO peaks were labeled with the name and retention time of the base peak. (<b>B</b>) The TIC from the colostrum samples on day 30 p.p. of the analyzed Holstein cattle were overlaid using FreeStyle software. The MO peaks were labeled with the name and retention time of the base peak. The ratios of the peak areas of the sialylated bMOs were determined (n = 5 animals), and box and whisker plots (median; min to max) are shown for (<b>C</b>) day 0 and (<b>D</b>) day 30 p.p. Only sialylated bMOs with peak areas greater than 0.1% were included. The statistical analysis and graphs were generated using BioRender.com. Significant differences are denoted as follows: *: <span class="html-italic">p</span> &lt; 0.05, **: <span class="html-italic">p</span> &lt; 0.01, ***: <span class="html-italic">p</span> &lt; 0.001, ****: <span class="html-italic">p</span> &lt; 0.0001. The bMO structures were designed with GlycoWorkbench 2 [<a href="#B43-foods-13-02484" class="html-bibr">43</a>].</p>
Full article ">Figure 8
<p>Abundance of the major sialylated bMOs during early lactation. The EICs were used to calculate the ratios of the peak areas of the most abundant sialylated bMOs. Box and whisker plots (median; min to max) are shown for (<b>A</b>) 3′-SL, (<b>B</b>) DSL, (<b>C</b>) 3′SLN, and (<b>D</b>) 6′-SL (n = 5 animals). Statistical analysis and graphs were generated using BioRender.com. Significant differences are denoted as follows: *: <span class="html-italic">p</span> &lt; 0.05, **: <span class="html-italic">p</span> &lt; 0.01, ***: <span class="html-italic">p</span> &lt; 0.001, ****: <span class="html-italic">p</span> &lt; 0.0001. The bMO structures were designed with GlycoWorkbench 2 [<a href="#B43-foods-13-02484" class="html-bibr">43</a>].</p>
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10 pages, 562 KiB  
Article
Presepsin in Human Milk Is Delivery Mode and Gender Dependent
by Ebe D’Adamo, Chiara Peila, Mariachiara Strozzi, Roberta Barolo, Antonio Maconi, Arianna Nanni, Valentina Botondi, Alessandra Coscia, Enrico Bertino, Francesca Gazzolo, Ali Saber Abdelhameed, Mariangela Conte, Simonetta Picone, Marianna D’Andrea, Mauro Lizzi, Maria Teresa Quarta and Diego Gazzolo
Nutrients 2024, 16(15), 2554; https://doi.org/10.3390/nu16152554 - 3 Aug 2024
Viewed by 746
Abstract
Breast milk (BM) is a unique food due to its nutritional composition and anti-inflammatory characteristics. Evidence has emerged on the role of Presepsin (PSEP) as a reliable marker of early sepsis diagnosis. In the present study, we aimed to investigate the measurability of [...] Read more.
Breast milk (BM) is a unique food due to its nutritional composition and anti-inflammatory characteristics. Evidence has emerged on the role of Presepsin (PSEP) as a reliable marker of early sepsis diagnosis. In the present study, we aimed to investigate the measurability of PSEP in BM according to different maturation stages (colostrum, C; transition, Tr; and mature milks, Mt) and corrected for delivery mode and gender. We conducted a multicenter prospective case–control study in women who had delivered 22 term (T) and 22 preterm (PT) infants. A total of 44 human milk samples were collected and stored at −80 °C. BM PSEP (pg/mL) levels were measured by using a rapid chemiluminescent enzyme immunoassay. PSEP was detected in all samples analyzed. Higher (p < 0.05) BM PSEP concentrations were observed in the PT compared to the T infants. According to the grade of maturation, higher (p < 0.05) levels of PSEP in C compared to Tr and Mt milks were observed in the whole study population. The BM subtypes’ degrees of maturation were delivery mode and gender dependent. We found that PSEP at high concentrations supports its antimicrobial action both in PT and T infants. These results open the door to further studies investigating the role of PSEP. Full article
(This article belongs to the Special Issue Breastfeeding: Benefits to Infant and Mother)
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Figure 1

Figure 1
<p>Flow chart describing patients’ recruitment. Abbreviations: PT, preterm; T, Term.</p>
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<p>Presepsin (PSEP) milk levels (pg/mL) measured in colostrum (C), transition (Tr) and mature (Mt) milk fluids in the preterm (PT) and term (T) groups. Values are expressed in median and interquartile ranges. <span class="html-italic">p</span> &lt; 0.05; * PSEP C vs. Tr vs. Mt; a PT PSEP C vs. Tr vs. Mt; b T PSEP C vs. Tr; &amp; PT PSEP C, Tr, Mt vs. T PSEP C, Tr, Mt.</p>
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14 pages, 503 KiB  
Article
Comparison of the Fatty Acid Profiles of Sow and Goat Colostrum
by Lucía Ayala, Pilar Gómez-Cortés, Fuensanta Hernández, Josefa Madrid, Silvia Martínez-Miró and Miguel Angel de la Fuente
Vet. Sci. 2024, 11(8), 341; https://doi.org/10.3390/vetsci11080341 - 29 Jul 2024
Viewed by 602
Abstract
Currently, the utilization of hyperprolific sows has stimulated the search for supplements aimed at enhancing piglet survival, as these sows yield more offspring than they can adequately feed with their colostrum production. In contrast, intensive goat farming often yields surplus colostrum, thus necessitating [...] Read more.
Currently, the utilization of hyperprolific sows has stimulated the search for supplements aimed at enhancing piglet survival, as these sows yield more offspring than they can adequately feed with their colostrum production. In contrast, intensive goat farming often yields surplus colostrum, thus necessitating its removal, since kids are exclusively fed colostrum through lactation solely within the initial day of birth. The objective of this study was to examine and compare the fatty acid (FA) profiles of colostrum from sows and goats, together with possible influencing factors such as sow parity and the postpartum day of the goat, for possible use as an energy supplement for neonatal piglets. Swine colostrum was collected from sows with a 0–5 parity. In addition, samples of goat colostrum were collected on their first (D1) and second (D2) days of postpartum milking. The FA profiles of the colostrum were analyzed via gas chromatography. The parity value of the sows did not affect (p > 0.05) the FA colostrum composition. High proportions of palmitic, oleic, and linoleic acids were found in both types of colostrum. Levels of palmitic, oleic, and linoleic acids were significantly higher in D1 goat colostrum, whereas saturated FAs of less than 14 carbons (4:0, 6:0, 8:0, 10:0, and 12:0) were found in higher proportions in D2. These FAs play an important role in colostrum as they are a readily available source of energy and have also been attributed strong antibacterial activity. Therefore, goat colostrum, especially D2, could be used as an alternative energy supplement for newborn piglets, in particular for the weakest and smallest of the litter, which are the most in need. Full article
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Figure 1

Figure 1
<p>Heatmap of the fatty acid composition of sow and goat colostrum collected on the first (D1) and second (D2) days of postpartum milking. The color of each tile is proportional to the normalized detected concentration; red indicates an increased value, whereas blue indicates a decreased value. Clustering was performed by calculating the Euclidean distance.</p>
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26 pages, 941 KiB  
Article
Changes in the Fatty Acid Profile of Lactating Women Living in Poland—A Comparison with the Fatty Acid Profile of Selected Infant Formulas
by Aleksandra Purkiewicz and Renata Pietrzak-Fiećko
Nutrients 2024, 16(15), 2411; https://doi.org/10.3390/nu16152411 - 25 Jul 2024
Viewed by 731
Abstract
The present study examined the fatty acid content of human milk from Polish women living in the Warmia and Mazury region with regard to different lactation periods and compared it with the fatty acid content of selected infant formulas. The analysis included samples [...] Read more.
The present study examined the fatty acid content of human milk from Polish women living in the Warmia and Mazury region with regard to different lactation periods and compared it with the fatty acid content of selected infant formulas. The analysis included samples of breast milk—colostrum (n = 21), transitional milk (n = 26), and mature milk (n = 22). Fat was extracted using the Rose-Gottlieb method, and the fatty acid profile was determined by gas chromatography with a flame ionization detector (FID). The proportion of SFAs (saturated fatty acids) > MUFAs (monounsaturated fatty acids) > PUFAs (polyunsaturated fatty acids) was determined in each fraction of breast milk and infant formula. Palmitic, oleic, and linoleic acids predominated in breast milk and infant formulas. Colostrum contained lower contents of selected SFAs (caprylic, capric, lauric) and higher contents of selected MUFAs (ercucic) and PUFAs (arachidonic and docosahexaenoic) (p < 0.05) relative to transitional and mature milk. Infant formulas were distinguished from human milk in terms of their SFA (caproic, caprylic, lauric, arachidic), MUFA (oleic), and PUFA (linoleic, α-linoleic) content. It should be noted that infant formulas contained significantly lower trans fatty acid (TFA) content—more than thirty-six and more than nineteen times lower than in human milk. Furthermore, human milk contained branched-chain fatty acids (BCFAs) at 0.23–0.28%, while infant formulas contained only trace amounts of these acids. The average ratio of n-6 to n-3 fatty acids for human milk was 6.59:1 and was close to the worldwide ratio of 6.53 ± 1.72:1. Both principal component analysis (PCA) and cluster analysis (CA) indicated significant differences in the fatty acid profile relative to lactation and a different profile of infant formulas relative to breast milk. Full article
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Figure 1

Figure 1
<p>The principal component plot showing variations in the selected fatty acids (C6:0, C8:0, C10:0, C12:0, C14:0, C15:0, C16:0, C17:0, C18:0, C20:0, C14:1 <span class="html-italic">n</span>-5, C16:1 <span class="html-italic">n</span>-7, C17:1 <span class="html-italic">n</span>-9, C18:1 <span class="html-italic">n</span>-9, C20:1 <span class="html-italic">n</span>-9, C22:1 <span class="html-italic">n</span>-9, C18:3 <span class="html-italic">n</span>-3, C20:5 <span class="html-italic">n</span>-3, C22:6 <span class="html-italic">n</span>-3, C18:2 <span class="html-italic">n</span>-6, C20:4 <span class="html-italic">n</span>-6, <span class="html-italic">iso</span> C15:0, <span class="html-italic">anteiso</span> C15:0, <span class="html-italic">iso</span> C16:0, C18:1 <span class="html-italic">n</span>6+<span class="html-italic">n</span>9 <span class="html-italic">t</span>, C18:1 <span class="html-italic">t</span>11, C18:2 <span class="html-italic">c</span>9<span class="html-italic">t</span>11, C16:1 <span class="html-italic">t</span>7) of the analyzed infant formulas and human milk samples (<b>A</b>) and the score plot of the analyzed human milk samples from different lactation periods and selected infant formulas (<b>B</b>). Explanations: C—colostrum, TM—transitional milk, MM—mature milk; IF-I, IF-II—infant formulas from different manufacturers.</p>
Full article ">Figure 2
<p>Cluster analysis (CA) of the fatty acid profile in studied human milk from different lactation periods and infant formulas. Abbreviations: IF-I, IF-II—infant formulas from different manufacturers.</p>
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21 pages, 4311 KiB  
Article
Biologically Active Sheep Colostrum for Topical Treatment and Skin Care
by Kinga Kazimierska, Ilona Szabłowska-Gadomska, Stefan Rudziński, Katarzyna Kośla, Elżbieta Płuciennik, Łukasz Bobak, Aleksandra Zambrowicz and Urszula Kalinowska-Lis
Int. J. Mol. Sci. 2024, 25(15), 8091; https://doi.org/10.3390/ijms25158091 - 25 Jul 2024
Viewed by 367
Abstract
Colostrum is gaining popularity in cosmetic products. The present study compared the composition and selected biological properties of colostrum from Polish sheep (colostrum 1) and Swiss sheep (colostrum 2), particularly those that can affect healthy or diseased skin. The antioxidant activity of the [...] Read more.
Colostrum is gaining popularity in cosmetic products. The present study compared the composition and selected biological properties of colostrum from Polish sheep (colostrum 1) and Swiss sheep (colostrum 2), particularly those that can affect healthy or diseased skin. The antioxidant activity of the colostrums was measured using ABTS and DPPH assays. The effect on the proliferation of human skin fibroblasts, neonatal epidermal keratinocytes, and human diabetic fibroblast (dHF) cells isolated from diabetic foot ulcers was also assayed in vitro by MTT and Presto Blue tests, respectively. The colostrum simulated dHF cell proliferation by up to 115.4%. The highest used concentration of colostrum 1 stimulated normal fibroblast proliferation by 191.2% (24 h) and 222.2% (48 h). Both colostrums inhibited epidermal keratinocyte viability. The influence of the colostrums on the expression of genes related to proliferation (Ki67) and immune response (IL-6, PTGS-2, TSG-6) in dHF cells were compared. Colostrum 1 increased the rate of wound closure (scar test). Analysis of total fat, protein and fatty acid content found the Polish colostrum to be a richer source of fat than the Swiss colostrum, which contained a larger amount of protein. Both colostrums exhibit properties that suggest they could be effective components in cosmetic or medicinal formulations for skin care, especially supporting its regeneration, rejuvenation, and wound healing. Full article
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Figure 1

Figure 1
<p>Percentage of ABTS radical inhibition by colostrum 1 (<b>a</b>) and colostrum 2 (<b>b</b>) over time (n = 3).</p>
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<p>The effect of colostrum 1 on the cell viability of: (<b>a</b>) 1BR.3.N human skin fibroblasts; (<b>b</b>) human neonatal epidermal keratinocytes (n = 6); *** <span class="html-italic">p</span> &lt; 0.001; ** <span class="html-italic">p</span> &lt; 0.01, * <span class="html-italic">p</span> &lt; 0.05.</p>
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<p>The effect of colostrum 2 on the cell viability of (<b>a</b>) 1BR.3.N human skin fibroblasts; (<b>b</b>) human neonatal epidermal keratinocytes (n = 6); *** <span class="html-italic">p</span> &lt; 0.001; ** <span class="html-italic">p</span> &lt; 0.01, * <span class="html-italic">p</span> &lt; 0.05.</p>
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<p>Presto Blue proliferation test results for diabetic human fibroblasts (dHF): (<b>a</b>) four different dHF cells samples (n = 3) cultured with either colostrum 1 or colostrum 2; (<b>b</b>) cumulative results of Presto Blue proliferation tests (n = 12). Proliferation rate was calculated with a dHF cultured in standard dHF medium as the control. The proliferation rate of stimulated dHF cells ranged from 96.0% (dHF1029—cultured with colostrum 1 after 24 h) to 115.4% (dHF1014 cultured with colostrum 2 after 48 h). SEMs were calculated. The statistical significance was calculated with Fisher’s test and Student’s <span class="html-italic">t</span>-test. *** <span class="html-italic">p</span> &lt; 0.001; ** <span class="html-italic">p</span> &lt; 0.01; * <span class="html-italic">p</span> &lt; 0.05.</p>
Full article ">Figure 4 Cont.
<p>Presto Blue proliferation test results for diabetic human fibroblasts (dHF): (<b>a</b>) four different dHF cells samples (n = 3) cultured with either colostrum 1 or colostrum 2; (<b>b</b>) cumulative results of Presto Blue proliferation tests (n = 12). Proliferation rate was calculated with a dHF cultured in standard dHF medium as the control. The proliferation rate of stimulated dHF cells ranged from 96.0% (dHF1029—cultured with colostrum 1 after 24 h) to 115.4% (dHF1014 cultured with colostrum 2 after 48 h). SEMs were calculated. The statistical significance was calculated with Fisher’s test and Student’s <span class="html-italic">t</span>-test. *** <span class="html-italic">p</span> &lt; 0.001; ** <span class="html-italic">p</span> &lt; 0.01; * <span class="html-italic">p</span> &lt; 0.05.</p>
Full article ">Figure 5
<p>Scar test results for dHF cells stimulated with either colostrum 1 or colostrum 2 (n = 4). (<b>a</b>) dHF cells stimulated with colostrum 1 closed the scar to 40% of the initial scar area (red line) after 18 h of culture and 20% (green line) after 28 h of culture. Colostrum 2 did not increase the speed of dHF migration compared to control culture. (<b>b</b>) Representative photographs of wounds in three selected timepoints. In the scar test, colostrum 1 had a stronger impact on scar closure rate. It was shown that ovine colostrum has the potential to enhance wound healing processes in diabetic foot ulcers.</p>
Full article ">Figure 5 Cont.
<p>Scar test results for dHF cells stimulated with either colostrum 1 or colostrum 2 (n = 4). (<b>a</b>) dHF cells stimulated with colostrum 1 closed the scar to 40% of the initial scar area (red line) after 18 h of culture and 20% (green line) after 28 h of culture. Colostrum 2 did not increase the speed of dHF migration compared to control culture. (<b>b</b>) Representative photographs of wounds in three selected timepoints. In the scar test, colostrum 1 had a stronger impact on scar closure rate. It was shown that ovine colostrum has the potential to enhance wound healing processes in diabetic foot ulcers.</p>
Full article ">Figure 6
<p>Expression of <span class="html-italic">Ki67</span>, <span class="html-italic">IL-6</span>, <span class="html-italic">PTGS-2</span>, and <span class="html-italic">TSG-6</span> in dHF after 48 h treatment with either colostrum 1 or colostrum 2 (n = 4). Panels: (<b>a</b>) dHF cells treated with colostrum 1 vs. control; (<b>b</b>) dHF cells treated with colostrum 2 vs. control; (<b>c</b>) colostrum 1 vs. colostrum 2. Statistical significance was determined in GraphPad program with the Mann–Whitney U-test with Holm–Šídák correction for multiple comparisons; **** <span class="html-italic">p</span> &lt; 0.0001; *** <span class="html-italic">p</span> &lt; 0.001; * <span class="html-italic">p</span> &lt; 0.05; ns means no significance.</p>
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15 pages, 1138 KiB  
Review
Revealing the Potency of Growth Factors in Bovine Colostrum
by Yalçın Mert Yalçıntaş, Hatice Duman, Jose M. Miranda López, Alicia C. Mondragón Portocarrero, Mauro Lombardo, Farid Khallouki, Wojciech Koch, Matteo Bordiga, Hesham El-Seedi, António Raposo, Jose Luiz de Brito Alves and Sercan Karav
Nutrients 2024, 16(14), 2359; https://doi.org/10.3390/nu16142359 - 21 Jul 2024
Viewed by 1038
Abstract
Colostrum is a nutritious milk synthesized by mammals during the postpartum period, and its rich bioactive components has led to a global increase in the consumption of bovine colostrum as a supplement. Bovine colostrum contains key components such as immunoglobulins, oligosaccharides, lactoferrin and [...] Read more.
Colostrum is a nutritious milk synthesized by mammals during the postpartum period, and its rich bioactive components has led to a global increase in the consumption of bovine colostrum as a supplement. Bovine colostrum contains key components such as immunoglobulins, oligosaccharides, lactoferrin and lysozyme. It is a special supplement source due to its natural, high bioavailability and high concentrations of growth factors. Growth factors are critical to many physiological functions, and considering its presence in the colostrum, further research must be conducted on its safe application in many bodily disorders. Growth factors contribute to wound healing, muscle and bone development, and supporting growth in children. Additionally, the molecular mechanisms have been explored, highlighting the growth factors roles in cell proliferation, tissue regeneration, and the regulation of immune responses. These findings are crucial for understanding the potential health effects of bovine colostrum, ensuring its safe use, and forming a basis for future clinical applications. This review article examines the growth factors concentration in bovine colostrum, their benefits, clinical studies, and molecular mechanisms. Full article
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<p>Growth factors in bovine colostrum [<a href="#B2-nutrients-16-02359" class="html-bibr">2</a>,<a href="#B9-nutrients-16-02359" class="html-bibr">9</a>].</p>
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<p>The molecular mechanism of the contribution of growth factors found in bovine colostrum to wound healing. Growth factors found in bovine colostrum stimulate the Ras/MAPK and PI3K/Akt signaling pathways through receptor binding, promoting the proliferation and differentiation of cells in the affected area. Consequently, the wound-healing process is accelerated [<a href="#B25-nutrients-16-02359" class="html-bibr">25</a>,<a href="#B26-nutrients-16-02359" class="html-bibr">26</a>]. (p85: regulatory subunit of phosphatidylinositol 3-kinase, p110: catalytic subunit of phosphatidylinositol 3-kinase, PTEN: phosphatase and tensin homolog, Akt: protein kinase B, PIP2: phosphatidylinositol (4,5)-bisphosphate, PIP3: phosphatidylinositol (3,4,5)-trisphosphate, TSC: tuberous sclerosis complex, Rheb: Ras homolog enriched in brain, mTORC1: mechanistic target of rapamycin complex, LKB1: liver kinase B1, AMPK1: AMP-activated protein kinase, GRB2: growth factor receptor-bound protein 2, SOS: Son of Sevenless, Ras: rat sarcoma, Raf: rapidly accelerated fibrosarcoma, MEK: mitogen-activated protein kinase, ERK: extracellular signal-regulated kinase).</p>
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13 pages, 695 KiB  
Article
Factors Influencing Breast Milk Antibody Titers during the Coronavirus Disease 2019 Pandemic: An Observational Study
by Christoph Hochmayr, Ira Winkler, Marlene Hammerl, Alexander Höller, Eva Huber, Martina Urbanek, Ursula Kiechl-Kohlendorfer, Elke Griesmaier and Anna Posod
Nutrients 2024, 16(14), 2320; https://doi.org/10.3390/nu16142320 - 18 Jul 2024
Viewed by 755
Abstract
The COVID-19 pandemic has highlighted the role of breastfeeding in providing passive immunity to infants via specific anti-SARS-CoV-2 antibodies in breast milk. We aimed to quantify these antibodies across different lactation stages and identify influencing factors. This prospective study involved mother–child dyads from [...] Read more.
The COVID-19 pandemic has highlighted the role of breastfeeding in providing passive immunity to infants via specific anti-SARS-CoV-2 antibodies in breast milk. We aimed to quantify these antibodies across different lactation stages and identify influencing factors. This prospective study involved mother–child dyads from Innsbruck University Hospital, Austria, with a positive maternal SARS-CoV-2 test during pregnancy or peripartum between 2020 and 2023. We collected breast milk samples at various lactation stages and analyzed anti-Spike S1 receptor-binding domain (S1RBD) immunoglobulins (Ig). Maternal and neonatal data were obtained from interviews and medical records. This study included 140 mothers and 144 neonates. Anti-S1RBD-IgA (72.0%), -IgG (86.0%), and -IgM (41.7%) were highly present in colostrum and decreased as milk matured. Mothers with natural infection and vaccination exhibited higher anti-S1RBD-IgA and -IgG titers in all milk stages. Mothers with moderate to severe infections had higher concentrations of anti-S1RBD-IgA and -IgG in transitional milk and higher anti-S1RBD-IgA and -IgM in mature milk compared to those with mild or asymptomatic infections. Variations in antibody responses were also observed with preterm birth and across different virus waves. This study demonstrates the dynamic nature of breast milk Ig and underscores the importance of breastfeeding during a pandemic. Full article
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<p>Concentrations of anti-SARS-CoV-2-S1RBD immunoglobulins (Ig) in different types of breast milk. This figure shows boxplots of anti-S1RBD-IgA, -IgG, and -IgM concentrations in various types of breast milk, including colostrum, transitional milk, and mature milk. Ig concentrations are plotted on the <span class="html-italic">y</span>-axis in U/mL. Center lines in boxes represent medians, box edges mark 1st and 3rd quartiles, and whiskers indicate 10th and 90th percentiles. Individual points indicate outliers. Crosses indicate mean Ig concentrations. The data in the box highlight the dynamic changes in breast milk Ig levels over the lactation period, with the highest median concentrations observed in colostrum and gradually decreasing median concentrations in transitional and mature milk. <span class="html-italic">Ig</span>, immunoglobulin; <span class="html-italic">SARS-CoV-2</span>, severe acute respiratory syndrome coronavirus 2; <span class="html-italic">S1RBD</span>, SARS-CoV-2 Spike protein S1 receptor-binding domain.</p>
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<p>Concentrations of anti-SARS-CoV-2-S1RBD immunoglobulins (Ig) in breast milk across different SARS-CoV-2 waves. This panel displays boxplots of anti-S1RBD-IgA, -IgG, and -IgM concentrations in different types of breast milk (colostrum, transitional milk, and mature milk) across several waves of the SARS-CoV-2 pandemic. Ig concentrations are plotted on the <span class="html-italic">y</span>-axis in U/mL. Center lines in boxes represent medians, box edges mark 1st and 3rd quartiles, and whiskers indicate 10th and 90th percentiles. The influence of different SARS-CoV-2 waves on breast milk Ig concentrations is depicted. Significant differences across different pandemic waves are indicated by asterisks (* <span class="html-italic">p</span> &lt; 0.05, ** <span class="html-italic">p</span> &lt; 0.01, *** <span class="html-italic">p</span> &lt; 0.001, **** <span class="html-italic">p</span> &lt; 0.0001). <span class="html-italic">Ig</span>, immunoglobulin; <span class="html-italic">SARS-CoV-2</span>, severe acute respiratory syndrome coronavirus 2; <span class="html-italic">S1RBD</span>, SARS-CoV-2 Spike protein S1 receptor-binding domain.</p>
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19 pages, 1017 KiB  
Article
Clostridium butyricum Probiotic Feed Additive: Modulation of Sow Milk Metabolomics and Mitigation of Pre-Weaning Piglet Diarrhea
by Jakavat Ruampatana, Junpen Suwimonteerabutr, Kunaporn Homyog, Wanwimon Mekboonsonglarp, Korntip Kanjanavaikoon, Wouter Van der Veken, Sutthasinee Poonyachoti, Takele Feyera, Sarn Settachaimongkon and Morakot Nuntapaitoon
Animals 2024, 14(14), 2098; https://doi.org/10.3390/ani14142098 - 18 Jul 2024
Viewed by 703
Abstract
The present study aimed to investigate the impact of Clostridium butyricum probiotic feed additive on sow and piglet performances, together with alterations in the lipidomic and metabolomic profiles of sow milk. Sixty-four Landrace × Yorkshire crossbred sows and 794 piglets were included. Sows [...] Read more.
The present study aimed to investigate the impact of Clostridium butyricum probiotic feed additive on sow and piglet performances, together with alterations in the lipidomic and metabolomic profiles of sow milk. Sixty-four Landrace × Yorkshire crossbred sows and 794 piglets were included. Sows were divided into two groups; i.e., (i) conventional gestation diet (control; n = 35) and (ii) conventional diet added with 10 g/sow/day of probiotic C. butyricum spores (treatment; n = 29) from one week before the estimated farrowing day until weaning (29.6 ± 4.8 days). The sow and piglet performances and incidence of piglet diarrhea were recorded. Changes in gross chemical composition, fatty acid and non-volatile polar metabolite profiles of sow colostrum, transient milk and mature milk were evaluated. The results showed that relative backfat loss in the treatment group (−2.3%) was significantly lower than in control group (11.6%), especially in primiparous sows (p = 0.019). The application of C. butyricum probiotics in sows significantly reduced the incidence of diarrhea in piglets (p < 0.001) but no other effect on piglet performance was found. Lipidomic and metabolomic analyses revealed variations in sow colostrum and milk biomolecular profiles, with indicative compounds significantly altered by feeding with the C. butyricum probiotics. In conclusion, the use of C. butyricum probiotics in sows may improve sow body condition and reduce diarrhea incidence in piglets, with underlying changes in milk composition that warrant further investigation. These findings support the potential of C. butyricum as a beneficial feed additive in swine production. Full article
(This article belongs to the Special Issue Feed Additives in Pig Feeding: 2nd Edition)
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<p>Incidence of diarrhea in piglets, from birth to 21 days in in the control and treatment groups. Level of significant difference at * <span class="html-italic">p</span> &lt; 0.05, ** <span class="html-italic">p</span> &lt; 0.01 and *** <span class="html-italic">p</span> &lt; 0.001.</p>
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<p>PLS-DA score plots for the comparison of fatty acid profiles of colostrum (day 0; panel (<b>A</b>)), transient milk (day 3; panel (<b>C</b>)) and mature milk (day 17; panel (<b>E</b>)) samples collected from sows in the control (red color) and <span class="html-italic">C. butyricum</span> treatment (blue color) groups. Samples from animals in parity 1 (■), parity 2–4 (▲) and parity 5–7 (●) are differently symbolized. VIP scores derived from the comparison among samples of the same day postpartum and indicative fatty acids accountable for the discrimination are visualized in panel (<b>B</b>), (<b>D</b>) and (<b>F</b>), respectively.</p>
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<p>PLS-DA score plots for the comparison of non-volatile polar metabolite profiles of colostrum (day 0; panel (<b>A</b>)), transient milk (day 3; panel (<b>C</b>)) and mature milk (day 17; panel (<b>D</b>)) samples collected from sows in the control (red color) and <span class="html-italic">C. butyricum</span> treatment (blue color) group. Samples from animals in parity 1 (■), parity 2–4 (▲) and parity 5–7 (●) are differently symbolized. VIP scores derived from the comparison among samples of the same day postpartum and indicative metabolites accountable for the discrimination are visualized in panel (<b>B</b>) and (<b>E</b>), respectively.</p>
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12 pages, 814 KiB  
Article
The Assessment of Iodine Concentrations in Colostrum and Breast Milk Using ICP-MS: The Impact of Delivery Type, Thyroid Function and Gestational Diabetes—A Pilot Study
by Jadwiga Kryczyk-Kozioł, Paulina Moniak, Paweł Zagrodzki, Ryszard Lauterbach, Hubert Huras, Magdalena Staśkiewicz, Mirosław Krośniak, Paweł Paśko, Robert Podsiadły and Justyna Dobrowolska-Iwanek
Foods 2024, 13(14), 2241; https://doi.org/10.3390/foods13142241 - 16 Jul 2024
Viewed by 492
Abstract
Considering the spectrum of benefits of breast milk feeding, determining the essential components of an infant’s only food-mother’s milk-seems justified, especially in the case of those whose deficiency (e.g., iodine) may result in developmental disorders. The main aim of this study was the [...] Read more.
Considering the spectrum of benefits of breast milk feeding, determining the essential components of an infant’s only food-mother’s milk-seems justified, especially in the case of those whose deficiency (e.g., iodine) may result in developmental disorders. The main aim of this study was the determination of the total iodine content of breast milk (including colostrum and mature milk). A secondary objective was to assess the influence of factors such as the type of delivery, hypothyroidism, gestational diabetes or the stage of lactation on this parameter. The study materials were colostrum and milk after 1 (n = 14), 2 and 3 months (n = 8) of lactation with a range of iodine concentrations (µg/L): 195–1648 and 170–842, 174–650 and 273–751, respectively. Iodine was determined using the inductively coupled plasma mass spectrometry (ICP-MS). Multivariate statistical analysis revealed, e.g., that delivery by caesarean section or dose of L-thyroxine taken by women to normalise thyroid hormones, had a significant effect on iodine concentrations in breast milk. Further research aimed at assessing the quality of breast milk should also include determining the factors influencing it. Full article
(This article belongs to the Special Issue Modern Analytical Methods for Food Ingredients)
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<p>The iodine concentration (µg/L) in colostrum (C) and milk samples collected from selected women over the course of the first three months of lactation (M1–M3), categorised by the mode of delivery: vaginally birth (<span class="html-italic">n</span> = 2; B1, A10) and caesarean section (<span class="html-italic">n</span> = 2; D3, D6).</p>
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<p>The coordinate values of the tested parameters in the system of the first two dimensions of CAM (meaning of symbols: 1—iodine concentration-quartile 1; 2—iodine concentration-quartile 2; 3—iodine concentration-quartile 3; 4—iodine concentration-quartile 4; 5—no hypothyroidism; 6—hypothyroidism; 7—no gestational diabetes mellitus; 8—gestational diabetes mellitus; 9—no L-thyroxine use; 10—L-thyroxine in a dose of less than 50 µg/day; 11—L-thyroxine at a dose of 50 to 100 µg/day; 12—L-thyroxine at a dose of more than 150 µg/day; 13—delivery by caesarean section; 14—vaginal delivery).</p>
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11 pages, 260 KiB  
Article
Effects of Dietary Lonicera flos and Sucutellaria baicalensis Mixed Extracts Supplementation on Reproductive Performance, Umbilical Cord Blood Parameters, Colostrum Ingredients and Immunoglobulin Contents of Late-Pregnant Sows
by Chengkun Fang, Xiaopeng Tang, Qingtai Zhang, Qifang Yu, Shengting Deng, Shusong Wu and Rejun Fang
Animals 2024, 14(14), 2054; https://doi.org/10.3390/ani14142054 - 12 Jul 2024
Viewed by 507
Abstract
The present study aimed to determine the effects of dietary Lonicera flos and Sucutellaria baicalensis mixed extract (LSE) supplementation during the late-pregnancy period on the reproductive performance, umbilical cord blood hematological parameters, umbilical cord serum biochemical parameters, immune indices, hormone levels, colostrum ingredients, [...] Read more.
The present study aimed to determine the effects of dietary Lonicera flos and Sucutellaria baicalensis mixed extract (LSE) supplementation during the late-pregnancy period on the reproductive performance, umbilical cord blood hematological parameters, umbilical cord serum biochemical parameters, immune indices, hormone levels, colostrum ingredients, and immunoglobulin contents of sows. A total of 40 hybrid pregnant sows were randomly assigned to the control group (CON; sows fed a basal diet) and LSE group (LSE; sows fed a basal diet supplemented with 500 g/t PE). The results indicated that dietary LSE supplementation significantly increased (p < 0.05) the number of alive and healthy piglets and the litter weight at birth, and significantly increased (p < 0.05) the platelet counts in umbilical cord blood. Dietary LSE supplementation significantly increased (p < 0.05) the levels of prolactin (PRL) and growth hormone (GH), and the content of interleukin 2 (IL-2) in umbilical cord serum. Moreover, immunoglobulin A (IgA) and immunoglobulin M (IgM) in the colostrum were increased with PE supplementation (p < 0.05). In conclusion, dietary LSE supplementation in late-pregnancy sows could improve reproductive performance and colostrum quality, and could also regulate the levels of reproductive hormone in umbilical cord serum. Full article
(This article belongs to the Special Issue Plant Extracts as Feed Additives in Animal Nutrition and Health)
19 pages, 4245 KiB  
Article
Analysis of Human Milk Microbiota in Northern Greece by Comparative 16S rRNA Sequencing vs. Local Dairy Animals
by Margaritis Tsifintaris, Michail Sitmalidis, Maria Tokamani, Christina Anastasiadi, Maria Georganta, Ilias Tsochantaridis, Dimitrios Vlachakis, Panagiotis Tsikouras, Nikolaos Nikolettos, George P. Chrousos, Raphael Sandaltzopoulos and Antonis Giannakakis
Nutrients 2024, 16(14), 2175; https://doi.org/10.3390/nu16142175 (registering DOI) - 9 Jul 2024
Viewed by 1308
Abstract
Milk is a biological fluid with a dynamic composition of micronutrients and bioactive molecules that serves as a vital nutrient source for infants. Milk composition is affected by multiple factors, including genetics, geographical location, environmental conditions, lactation phase, and maternal nutrition, and plays [...] Read more.
Milk is a biological fluid with a dynamic composition of micronutrients and bioactive molecules that serves as a vital nutrient source for infants. Milk composition is affected by multiple factors, including genetics, geographical location, environmental conditions, lactation phase, and maternal nutrition, and plays a key role in dictating its microbiome. This study addresses a less-explored aspect, comparing the microbial communities in human breast milk with those in mature milk from species that are used for milk consumption. Since mature animal milk is used as a supplement for both the infant (formula) and the child/adolescent, our main aim was to identify shared microbial communities in colostrum and mature human milk. Using 16S rRNA metagenomic sequencing, we focused on characterizing the milk microbiota in the Northern Greek population by identifying shared microbial communities across samples and comparing the relative abundance of prevalent genera. We analyzed ten human milk samples (from five mothers), with five collected three days postpartum (colostrum) and five collected thirty to forty days postpartum (mature milk) from corresponding mothers. To perform an interspecies comparison of human milk microbiota, we analyzed five goat and five bovine milk samples from a local dairy industry, collected fifty to seventy days after birth. Alpha diversity analysis indicated moderate diversity and stability in bovine milk, high richness in goat milk, and constrained diversity in breast milk. Beta diversity analysis revealed significant distinctions among mammalian species, emphasizing both presence/absence and abundance-based clustering. Despite noticeable differences, shared microbial components underscore fundamental aspects across all mammalian species, highlighting the presence of a core microbiota predominantly comprising the Proteobacteria, Firmicutes, and Actinobacteriota phyla. At the genus level, Acinetobacter, Gemella, and Sphingobium exhibit significant higher abundance in human milk compared to bovine and goat milk, while Pseudomonas and Atopostipes are more prevalent in animal milk. Our comparative analysis revealed differences and commonalities in the microbial communities of various mammalian milks and unraveled the existence of a common fundamental milk core microbiome. We thus revealed both species-specific and conserved microbial communities in human, bovine, and goat milk. The existence of a common core microbiome with conserved differences between colostrum and mature human milk underscores fundamental similarities in the microbiota of milk across mammalian species, which could offer valuable implications for optimizing the nutritional quality and safety of dairy products as well as supplements for infant health. Full article
(This article belongs to the Special Issue Breastmilk for Healthy Development)
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<p>Bovine, goat, and human mature milk microbiome alpha diversity. (<b>A</b>) Chao1, (<b>B</b>) Simpson, and (<b>C</b>) Shannon indices and (<b>D</b>) Pielou’s evenness. Statistical significance (Wilcoxon test); NS <span class="html-italic">p</span> &gt; 0.05, (*) <span class="html-italic">p</span> &lt; 0.05, (**) <span class="html-italic">p</span> &lt; 0.001.</p>
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<p>Principal coordinates analysis (PCoA) plot based on the (<b>A</b>) Bray-Curtis dissimilarity matrix (<b>B</b>) Jaccard index, (<b>C</b>) unweighted UniFrac distance, and (<b>D</b>) weighted UniFrac distance matrix, calculated for genus-level abundances and depicting patterns of beta diversity for microbial communities in milk.</p>
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<p>Taxonomic summary of amplicon sequence variants detected in milk samples. (<b>A</b>) Total number of ASVs at the phylum, order, and genus levels. (<b>B</b>) A stacked bar graph shows the relative abundance of microorganisms detected in milk samples. Relative frequencies smaller than 1% were collapsed into the “other” category. Human-A: colostrum milk; Human-B: mature milk.</p>
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<p>Comparison of core microbiota in bovine, goat, and human milk (colostrum and mature) at the genus level. (<b>A</b>) Venn diagram illustrating the numbers of shared and unique amplicon sequence variants between three species. (<b>B</b>) Sankey plot depicting the genera that are detected consistently in human breast milk samples in timepoint A (human-A: colostrum milk) and timepoint B (human-B: mature milk). (<b>C</b>) Radial network graph depicting genus-level overlap of bacterial communities in the milk microbiota of bovine, goat, and human milk. The detection rate threshold was set to 80% (4/5 samples).</p>
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<p>Comparative analysis of core milk bacterial abundance between humans and local dairy animals. Bacterial genera that met the detection rate threshold (80%; detected in at least four out of five samples) are depicted with green dots, whereas those that did not meet the threshold are indicated with red dots. Differential abundance results for genera that did not meet the detection threshold were not considered and are marked with a dash (“-”). Results with an adjusted <span class="html-italic">p</span>-value less than 0.05 were considered statistically significant (four stars: <span class="html-italic">p</span> &lt; 0.0001, three stars: <span class="html-italic">p</span> &lt; 0.001, two stars: <span class="html-italic">p</span> &lt; 0.01, one star: <span class="html-italic">p</span> &lt; 0.05, no stars: <span class="html-italic">p</span> ≥ 0.05). Relative abundance rank is indicated by color: high (1–60) in black, medium (61–120) in red, and low (&gt;120) in green.</p>
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15 pages, 3106 KiB  
Article
Bacillus subtilis Feed Supplementation Combined with Oral E. coli Immunization in Sows as a Tool to Reduce Neonatal Diarrhea in Piglets
by Jianxin Liu, Danchen Aaron Yang, Haobo Qu, Dandan Liu and Kehe Huang
Animals 2024, 14(13), 1978; https://doi.org/10.3390/ani14131978 - 4 Jul 2024
Viewed by 555
Abstract
To investigate the effects of B. subtilis on the specific immune response of lactating sows to E. coli and the diarrhea rate in suckling piglets, thirty large white sows with similar farrowing dates were randomly divided into two groups: a feedback feeding (i.e., [...] Read more.
To investigate the effects of B. subtilis on the specific immune response of lactating sows to E. coli and the diarrhea rate in suckling piglets, thirty large white sows with similar farrowing dates were randomly divided into two groups: a feedback feeding (i.e., feeding a homogenate of intestinal contents and tissues from E. coli-infected piglets to sows; FB) group and a feedback feeding with B. subtilis (FB + BS) group. Serum, colostrum, and intestinal tissues from sows and piglets were collected to assess the immune response and intestinal barrier function at weaning. T and B cells from Peyer’s patches (PPs) and mesenteric lymph nodes (MLNs) in lactating mice (with treatments consistent with the sows’) were isolated to explore the underlying mechanism. The results showed that, compared with the FB group, the reproductive performance of sows and the growth performance of their offspring were effectively improved in the FB + BS group. Moreover, the levels of IgG/IgA and those of IgG/IgA against E. coli in the serum and colostrum of sows in the FB+BS group were increased (p < 0.05). Meanwhile, the ratio of CD4+/CD8+, CD4+CXCR5+PD1+, and B220+IgA+ cells in MLNs and PPs, and the IgA levels in the mammary glands of mice, were also increased in the FB + BS group (p < 0.05). Notably, in suckling piglets in the FB + BS group, the diarrhea rate was decreased (p < 0.05), and the intestinal barrier function and intestinal flora composition at weaning were significantly improved. Overall, these results indicated that B. subtilis feed supplementation combined with feedback feeding in pregnant and lactating sows can reduce diarrhea in suckling piglets by enhancing the maternal immune response against E. coli and intestinal barrier function in their offspring, improving survival rates and pre-weaning growth. Full article
(This article belongs to the Section Pigs)
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<p>Effects of <span class="html-italic">B. subtilis</span> on the serum and colostrum antibody levels of sows after the <span class="html-italic">E. coli</span> challenge. (<b>A</b>) The concentrations of IgA in serum (geometric mean) were measured by ELISA; (<b>B</b>) The concentrations of IgG (geometric mean) in serum were measured by ELISA; (<b>C</b>) The concentrations of F4ac-IgG and F4ac-IgA in serum were measured by ELISA; (<b>D</b>) The concentrations of IgG and IgA in colostrum were measured by ELISA; (<b>E</b>) The concentrations of F4ac-IgG and F4ac-IgA in colostrum were measured by ELISA. Data are shown as medians (IQR), N = 5. * <span class="html-italic">p</span> &lt; 0.05 and ** <span class="html-italic">p</span> &lt; 0.01 were considered statistically significant.</p>
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<p>Effects of <span class="html-italic">B. subtilis</span> on the antibody levels and IgA-producing plasma cell accumulation of mice after the <span class="html-italic">E. coli</span> challenge. (<b>A</b>) Concentrations of IgG and IgA in serum and SIgA in intestinal mucus were measured by ELISA; (<b>B</b>) MLN and PP representative flow cytometry plots (including percentages) of CD4+CD8+ cells, CD4+CXCR5+PD1+ cells, and B220+IgA+ cells; (<b>C</b>) H&amp;E staining of the mammary tissue (scale bar = 500 μm); (<b>D</b>) Immunohistochemical diagram of IgA in mammary tissue (scale bar = 500 μm). Data are shown as medians (IQR), N = 5. * <span class="html-italic">p</span> &lt; 0.05 and ** <span class="html-italic">p</span> &lt; 0.01 were considered statistically significant.</p>
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<p>Effects of <span class="html-italic">B. subtilis</span> on serum antibody levels and intestinal barrier function of piglets born to <span class="html-italic">E. coli</span>-challenged sows. (<b>A</b>) The concentrations of IgG and IgA in serum were measured by ELISA; (<b>B</b>) The mRNA levels of <span class="html-italic">ZO</span>-<span class="html-italic">1</span>, <span class="html-italic">occludin</span>, <span class="html-italic">claudin1</span>, and <span class="html-italic">muc</span>-<span class="html-italic">2</span> were measured by RT-qPCR; (<b>C</b>) The immunofluorescence diagram of <span class="html-italic">ZO</span>-<span class="html-italic">1</span> in intestinal tissue, (scale bar = 200 μm); (<b>D</b>) The H&amp;E staining of the intestinal tissue, (scale bar = 200 μm). The data are shown as median (IQR), N = 5. * <span class="html-italic">p</span> &lt; 0.05 and ** <span class="html-italic">p</span> &lt; 0.01 were considered statistically significant.</p>
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<p>Effects of <span class="html-italic">B. subtilis</span> on the gut microbiota diversity of suckling piglets born to <span class="html-italic">E. coli</span>-challenged sows. (<b>A</b>) Fecal microbial alpha diversity at the species level was estimated by the Chao1 index, Shannon index, and Simpson index; (<b>B</b>) PCA and PCoA analysis of piglet rectum content microbiome; (<b>C</b>) Venn diagram displaying group overlaps; (<b>D</b>) Average relative abundance of the dominant phylum and genus in each group. Data are shown as median (IQR), N = 5. * <span class="html-italic">p</span> &lt; 0.05 was considered statistically significant.</p>
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15 pages, 2049 KiB  
Article
Feasibility Trial Exploring Immune-Related Biomarkers Pertaining to Rapid Immune Surveillance and Cytokine Changes after Consuming a Nutraceutical Supplement Containing Colostrum- and Egg-Based Low-Molecular-Weight Peptides
by Liu Yu, Ifeanyi Iloba, Dina Cruickshank and Gitte S. Jensen
Curr. Issues Mol. Biol. 2024, 46(7), 6710-6724; https://doi.org/10.3390/cimb46070400 - 30 Jun 2024
Viewed by 732
Abstract
Immune protection associated with consuming colostrum-based peptides is effective against bacterial and viral insults. The goal for this study was to document acute changes to immune surveillance and cytokine levels after consuming a single dose of a nutraceutical blend in the absence of [...] Read more.
Immune protection associated with consuming colostrum-based peptides is effective against bacterial and viral insults. The goal for this study was to document acute changes to immune surveillance and cytokine levels after consuming a single dose of a nutraceutical blend in the absence of an immune challenge. A double-blind, randomized, placebo-controlled, cross-over pilot study involved healthy participants attending two clinic visits. Blood draws were performed pre-consumption and at 1, 2, and 24 h after consuming a blend of bovine colostrum- and hen’s egg-based low-molecular-weight peptides (CELMPs) versus a placebo. Immunophenotyping was performed by flow cytometry, and serum cytokines were measured by multiplex cytokine arrays. Consumption of CELMPs triggered increased immune surveillance after 1 h, involving monocytes (p < 0.1), natural killer (NK) cells (p < 0.1), and natural killer T (NKT) cells (p < 0.05). The number of NKT cells expressing the CD25 immunoregulatory marker increased at 1 and 2 h (p < 0.1). Increased serum levels of monocyte chemoattractant protein-1 (MCP-1) was observed at 2 and 24 h (24 h: p < 0.05). Selective reduction in pro-inflammatory cytokines was seen at 1, 2, and 24 h, where the 2-h reduction was highly significant for IL-6, IFN-γ, and IL-13. The rapid, transient increase in immune surveillance, in conjunction with the reduced levels of inflammatory markers, suggests that the CELMP blend of natural peptides provides immune benefits of use in preventive medicine. Further studies are warranted in chronic inflammatory conditions. Full article
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<p>Diagram showing the involvement where all study participants were tested on two different clinic days, with blood draws at the baseline (T0), 1 h (T1), 2 h (T2), and 24 h (T24) after consuming a test product. An additional blood draw was performed 24 h after product consumption for both clinic days. The red arrows represent blood draw times as part of the core study design, at the baseline (T0), 1 h (T1), and 2 h (T2), and at 24 h after consuming a test product. For each sample collection time point, saliva was also collected (passive drool).</p>
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<p>Differences in changes to immune cell trafficking reflected by the numbers of immune cells in the blood circulation within 2 h after consumption of CELMPs versus the placebo. The results are shown as the group averages ± standard error of the mean of the individual percent changes from the baseline after consuming CELMPs, where changes after consuming the placebo are subtracted. (<b>A</b>) Monocyte numbers: A gradual reduction was seen at 1 and 2 h, and the reduction at 1 h reached a statistical trend. (<b>B</b>) NK cell numbers: An increase was seen at 1 h, returning to similar levels as after consuming the placebo at 2 h. The increase at 1 h reached a statistical trend. (<b>C</b>) NKT cell numbers: An increase after consuming the CELMPs compared to the placebo reached statistical significance at 1 h, returning to similar levels to after consuming the placebo at 2 h. (<b>D</b>) Numbers of T cells: No significant differences were observed in the T cell numbers after consuming CELMPs compared to the placebo. Levels of statistical significance are shown in the graphs, where changes from the baseline to a later time point are indicated by asterisks: <span class="html-italic">p</span> &lt; 0.10: (*), <span class="html-italic">p</span> &lt; 0.05: *.</p>
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<p>Differences in changes to expression of the immunoregulatory marker CD25 on 4 types of immune cells in the blood circulation within 2 h after consumption of CELMPs versus the placebo. The results are shown as the group averages ± standard error of the mean of the individual percent changes from the baseline after consuming CELMPs, where changes after consuming the placebo are subtracted. (<b>A</b>) CD25 expression on monocytes: A gradual increase was seen at 1 and 2 h, not reaching statistical significance. (<b>B</b>) CD25 expression on NK cells: A gradual increase was seen at 1 and 2 h, reaching a statistical trend at 2 h. (<b>C</b>) CD25 expression on NKT cells: An increase after consuming the CELMPs compared to the placebo reached statistical trends at 1 and 2 h. (<b>D</b>) CD25 expression on T cells: No significant differences were observed after consuming CELMPs compared to the placebo. Levels of statistical significance are shown in the graphs, where changes from the baseline to a later time point are indicated by asterisk: <span class="html-italic">p</span> &lt; 0.10: (*).</p>
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<p>Differences in changes to expression of CD69 on 4 types of immune cells in the blood circulation within 2 h after consumption of CELMPs versus the placebo. The results are shown as the group averages ± standard error of the mean of the individual percent changes from the baseline after consuming CELMPs, where changes after consuming the placebo are subtracted. (<b>A</b>) CD69 expression on monocytes: A gradual increase was seen at 1 h, reaching statistical trend and returned to similar levels to after consuming the placebo at 2 h. (<b>B</b>) CD69 expression on NK cells: No significant differences were observed after consuming CELMPs compared to the placebo. (<b>C</b>) CD69 expression on NKT cells: No significant differences were observed after consuming CELMPs compared to the placebo. (<b>D</b>) CD69 expression on T cells: No significant differences were observed after consuming CELMPs compared to the placebo. Levels of statistical significance are shown in the graphs, where changes from the baseline to a later time point are indicated by asterisk: <span class="html-italic">p</span> &lt; 0.10: (*).</p>
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<p>Differences in changes to cytokine levels in the blood circulation within 2 h after consumption of CELMPs versus the placebo. The results are shown as the group averages ± standard error of the mean of the individual percent changes from the baseline after consuming CELMPs, where changes after consuming the placebo are subtracted. (<b>A</b>) Interferon-gamma (IFN-γ) levels: A gradual decrease was seen at 1 and 2 h, reaching a statistical trend at 1 h and a high level of significance at 2 h. (<b>B</b>) Interleukin-6 (IL-6) levels: A gradual decrease was seen at 1 and 2 h, reaching a statistical trend at 1 h and a high level of significance at 2 h. (<b>C</b>) Interleukin-13 (IL-13) levels: A gradual decrease was seen at 2 h, reaching a high level of significance at 2 h. (<b>D</b>) Interleukin-17 (IL-17) levels: No significant differences were observed after consuming CELMPs compared to the placebo. (<b>E</b>) Monocyte chemoattractant protein-1 (MCP-1): An increase was observed at 2 h but did not reach statistical significance. (<b>F</b>) Tumor necrosis factor-alpha (TNF-α) levels: A gradual decrease was seen at 2 h but did not reach statistical significance. Levels of statistical significance are shown in the graphs, where changes from the baseline to a later time point are indicated by asterisks: <span class="html-italic">p</span> &lt; 0.10: (*) and <span class="html-italic">p</span> &lt; 0.01: **.</p>
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<p>Differences in changes to immune markers in the blood circulation 24 h after consumption of CELMPs versus the placebo. The results are shown as the group averages ± standard error of the mean of the individual percent changes from the baseline after consuming CELMPs, where changes after consuming the placebo are subtracted. (<b>A</b>) The numbers of monocytes, NK cells, and NKT cells increased, whereas there were no changes to T cell numbers. The increased numbers of monocytes reached a statistical trend, the numbers of NK cells were significant, and the increased numbers of NKT cells were highly significant. (<b>B</b>) Cytokine levels: Four cytokines showed reduced levels: interferon-gamma (IFN-γ), interleukin-6 (IL-6), interleukin-13 (IL-13), and tumor necrosis factor-alpha (TNF-α). The reduced levels of IFN-γ and IL-6 reached a statistical trend, and the reduced level of IL-6 was mild but statistically significant. No change was seen for IL-17. In contrast, a statistically significant increase in monocyte chemoattractant protein-1 (MCP-1) was observed at 24 h after consumption. Levels of statistical significance in changes from the baseline to a later time point are indicated by asterisks: <span class="html-italic">p</span> &lt; 0.10: (*), <span class="html-italic">p</span> &lt; 0.05: *, and <span class="html-italic">p</span> &lt; 0.01: **.</p>
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17 pages, 3035 KiB  
Article
A New Method for Growth Factor Enrichment from Dairy Products by Electrodialysis with Filtration Membranes: The Major Impact of Raw Product Pretreatment
by Sabita Kadel, Vladlen Nichka, Jacinthe Thibodeau, Behnaz Razi Parjikolaei and Laurent Bazinet
Int. J. Mol. Sci. 2024, 25(13), 7211; https://doi.org/10.3390/ijms25137211 - 29 Jun 2024
Viewed by 594
Abstract
This study is focused on fractionation of insulin-like growth factor I (IGF-I) and transforming growth factor-β2 (TGF-β2) using a new electro-based membrane process calledelectrodialysis with filtration membranes (EDFM). Before EDFM, different pretreatments were tested, and four pH conditions (4.25, 3.85, 3.45, and 3.05) [...] Read more.
This study is focused on fractionation of insulin-like growth factor I (IGF-I) and transforming growth factor-β2 (TGF-β2) using a new electro-based membrane process calledelectrodialysis with filtration membranes (EDFM). Before EDFM, different pretreatments were tested, and four pH conditions (4.25, 3.85, 3.45, and 3.05) were used during EDFM. It was demonstrated that a 1:1 dilution of defatted colostrum with deionized water to decrease mineral content followed by the preconcentration of GFs by UF is necessary and allow for these compounds to migrate to the recovery compartment during EDFM. MS analyses confirmed the migration, in low quantity, of only α-lactalbumin (α-la) and β-lactoglobulin (β-lg) from serocolostrum to the recovery compartment during EDFM. Consequently, the ratio of GFs to total protein in recovery compartment compared to that of feed serocolostrum solution was 60× higher at pH value 3.05, the optimal pH favoring the migration of IGF-I and TGF-β2. Finally, these optimal conditions were tested on acid whey to also demonstrate the feasibility of the proposed process on one of the main by-products of the cheese industry; the ratio of GFs to total protein was 2.7× higher in recovery compartment than in feed acid whey solution, and only α-la migrated. The technology of GF enrichment for different dairy solutions by combining ultrafiltration and electrodialysis technologies was proposed for the first time. Full article
(This article belongs to the Special Issue Feature Papers in 'Physical Chemistry and Chemical Physics' 2024)
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<p>Total protein concentration in recovery compartment after 180 min of EDFM experiment for different pH conditions. Data with different letters are significantly different.</p>
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<p>MS chromatogram of serocolostrum 2.38× in feed (T0) and cationic fractions (recovered after 180 min of EDFM experiment).</p>
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<p>The concentration of IGF-I and TGF-β2 in the recovery compartment after 180 min of the EDFM experiment. Data with different letters are significantly different. Lowercase letters indicate differences for IGF-I concentration; uppercase—for TGF-β2 concentration.</p>
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<p>The enrichment of the GFs of interest (IGF-I+TGF-β2) in the recovery compartment at T180. Data with different letters are significantly different.</p>
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<p>The evolution of relative energy consumption as a function of the total concentration of GFs in the recovery compartment after 180 min of EDFM. Data with different letters are significantly different.</p>
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<p>The concentration of TGF-β2 and IGF-I in the (<b>a</b>) feed and (<b>b</b>) recovery compartments during 180 min of EDFM of whey. Data with different letters are significantly different. Lowercase letters indicate differences for IGF-I concentration; uppercase—for TGF-β2 concentration.</p>
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<p>Enrichment in (<b>a</b>) the GFs of interest (IGF-I+TGF-β2) and (<b>b</b>) α-lactalbumin in the feed (at T0 and T180) and the recovery compartment at T180. Data with different letters are significantly different.</p>
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<p>EDFM configuration, consisting of one recovery compartment, to produce GF-enriched fractions. AEM: Anion exchange membrane; UFM: Ultrafiltration membrane; CEM: Cation exchange membrane; P<sup>+</sup>: Positively charged protein; P<sup>−</sup>: Negatively charged protein; P<sup>0</sup>: Neutral protein.</p>
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12 pages, 2330 KiB  
Article
Human Milk Protein-Derived Bioactive Peptides from In Vitro-Digested Colostrum Exert Antimicrobial Activities against Common Neonatal Pathogens
by Yang Lyu, Bum Jin Kim, Jagdish Suresh Patel, David C. Dallas and Yimin Chen
Nutrients 2024, 16(13), 2040; https://doi.org/10.3390/nu16132040 - 27 Jun 2024
Viewed by 840
Abstract
Human milk reduces risk for necrotizing enterocolitis in preterm infants. Necrotizing enterocolitis occurs in the ileocecal region where thousands of milk protein-derived peptides have been released from digestion. Digestion-released peptides may exert bioactivity, such as antimicrobial and immunomodulatory activities, in the gut. In [...] Read more.
Human milk reduces risk for necrotizing enterocolitis in preterm infants. Necrotizing enterocolitis occurs in the ileocecal region where thousands of milk protein-derived peptides have been released from digestion. Digestion-released peptides may exert bioactivity, such as antimicrobial and immunomodulatory activities, in the gut. In this study, we applied mass spectrometry-based peptidomics to characterize peptides present in colostrum before and after in vitro digestion. Sequence-based computational modeling was applied to predict peptides with antimicrobial activity. We identified more peptides in undigested samples, yet the abundances were much higher in the digested samples. Heatmapping demonstrated highly different peptide profiles between undigested and digested samples. Four peptides (αS1-casein [157–163], αS1-casein [157–165], β-casein [153–159] and plasminogen [591–597]) were selected, synthesized and tested against common pathogenic bacteria associated with necrotizing enterocolitis. All four exhibited bacteriostatic, though not bactericidal, activities against Klebsiella aerogenes, Citrobacter freundii and Serratia marcescens, but not Escherichia coli. Full article
(This article belongs to the Special Issue Bioactive Milk Proteins and Human Health)
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<p>Venn diagram and bar graph of (<b>a</b>) counts; (<b>b</b>) total chromatographic abundances (Log10) of human milk peptides identified from undigested and digested human milk samples; (<b>c</b>) Pie charts show the percentage of the total abundance of common (16% and 17%, respectively, in dark grey) and unique (84% and 83%, respectively) proteins identified in undigested (in white) and digested (in black) samples. * <span class="html-italic">p</span> = 0.005; difference in total peptide abundance between undigested and digested using related-samples Wilcoxon Signed Rank test.</p>
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<p>Top 10 parent proteins based on highest number of peptides that were derived in both undigested (blue) and digested (red) human milk samples.</p>
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<p>Heatmap and hierarchical clustering showing the similarity/differences of abundance pattern of human milk proteins that were identified using peptide profiles between undigested (U) and digested (D) samples.</p>
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<p>Percent change in optical density at 600 nm from the growth of common pathogenic bacteria incubated with synthesized peptides SP1-SP4. (<b>a</b>) <span class="html-italic">C. freundii</span>; (<b>b</b>) <span class="html-italic">E. coli</span>; (<b>c</b>) <span class="html-italic">K. aerogenes</span>; (<b>d</b>) <span class="html-italic">S. marcescens.</span></p>
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