Frozen Mother’s Own Milk Can Be Used Effectively to Personalize Donor Human Milk

Feeding preterm infants mother’s own milk (MOM) lowers rates of sepsis, decreases necrotizing enterocolitis, and shortens hospital stay. In the absence of freshly expressed MOM, frozen MOM (FMOM) is provided. When MOM is unavailable, preterm infants are often fed pasteurized donor human milk (DHM), rendering it devoid of beneficial bacteria. We have previously reported that when MOM is inoculated into DHM to restore the live microbiota [restored milk (RM)], a similar microbial diversity to MOM can be achieved. Yet, it is unknown if a similar diversity to MOM can be obtained when FMOM is inoculated into DHM. The goal of this study was to determine whether a similar microbial composition to MOM could be obtained when FMOM is used to personalize DHM. To this end, a fresh sample of MOM was obtained and divided into fresh and frozen fractions. MOM and FMOM were inoculated into DHM at different dilutions: MOM/FMOM 10% (RM/FRM10) and MOM/FMOM 30% (RM/FRM30) and incubated at 37°C. At different timepoints, culture-dependent and culture-independent techniques were performed. Similar microbiota expansion and alpha diversity were observed in MOM, RM10, and RM30 whether fresh or frozen milk was used as the inoculum. To evaluate if microbial expansion would result in an abnormal activation on the innate immune system, Caco-2 epithelial cells were exposed to RM/FRM to compare interleukin 8 levels with Caco-2 cells exposed to MOM or DHM. It was found that RM samples did not elicit a significant increase in IL-8 levels when compared to MOM or FMOM. These results suggest that FMOM can be used to inoculate DHM if fresh MOM is unavailable or limited in supply, allowing both fresh MOM and FMOM to be viable options in a microbial restoration strategy.

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Inoculation of mother’s own milk could personalize pasteurized donor human milk used for feeding preterm infants

Journal of Translational Medicine ◽

10.1186/s12967-021-03096-7 ◽

2021 ◽

Vol 19 (1) ◽

Author(s):

D. Mallardi ◽

C. Tabasso ◽

P. Piemontese ◽

S. Morandi ◽

T. Silvetti ◽

...

Keyword(s):

Human Milk ◽

Preterm Infants ◽

Bacterial Growth ◽

Microbiological Quality ◽

In Vitro Study ◽

Total Bacterial Count ◽

Milk Samples ◽

Donor Human Milk ◽

Mother’S Own Milk

Abstract Background Human milk is a vehicle for bioactive compounds and beneficial bacteria which promote the establishment of a healthy gut microbiome of newborns, especially of preterm infants. Pasteurized donor human milk (PDHM) is the second-best option when preterm mother’s own milk is unavailable. Since pasteurization affect the microbiological quality of donor milk, PDHM was inoculated with different preterm milk samples and then incubated, in order to evaluate the effect in terms of bacterial growth, human milk microbiome and proteolytic phenomena. Methods In an in-vitro study PDHM was inoculated at 10% v/v using ten preterm milk samples. Microbiological, metataxonomic and peptidomic analyses, on preterm milk samples at the baseline (T0), on PDHM and on inoculated milk (IM) samples at T0, after 2 h (T1) and 4 h (T2) of incubation at 37 °C, were conducted. Results IM samples at T2 showed a Total Bacterial Count not significantly different (p > 0.01) compared to preterm milk samples. At T2 lactic acid bacteria level was restored in all IM. After inoculation, metataxonomic analysis in IM samples showed that Proteobacteria remained the predominant phylum while Firmicutes moved from 3% at T1 to 9.4% at T2. Peptidomic profile of IM resembled that of PDHM, incubated for the same time, in terms of number and type of peptides. Conclusion The study demonstrated that inoculation of PDHM with mother’s own milk could restore bacterial growth and personalize human milk microbiome in PDHM. This effect could be beneficial because of the presence of maternal probiotic bacteria which make PDHM more similar to mother’s own milk.

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Metabolomic Profile of Personalized Donor Human Milk

Molecules ◽

10.3390/molecules25245783 ◽

2020 ◽

Vol 25 (24) ◽

pp. 5783

Author(s):

Monica F. Torrez Lamberti ◽

Evon DeBose-Scarlett ◽

Timothy Garret ◽

Leslie Ann Parker ◽

Josef Neu ◽

...

Keyword(s):

Human Milk ◽

Complex Mixture ◽

Metabolomic Analysis ◽

Metabolomic Profile ◽

Beneficial Bacteria ◽

Donor Human Milk ◽

Metabolite Profiles ◽

Metabolite Concentrations ◽

Mother’S Own Milk ◽

The Impact

Human milk could be considered an active and complex mixture of beneficial bacteria and bioactive compounds. Since pasteurization drastically reduces the microbial content, we recently demonstrated that pasteurized donor human milk (DHM) could be inoculated with different percentages (10% and 30%) of mother’s own milk (MOM) to restore the unique live microbiota, resulting in personalized milk (RM10 and RM30, respectively). Pasteurization affects not only the survival of the microbiota but also the concentration of proteins and metabolites, in this study, we performed a comparative metabolomic analysis of the RM10, RM30, MOM and DHM samples to evaluate the impact of microbial restoration on metabolite profiles, where metabolite profiles clustered into four well-defined groups. Comparative analyses of DHM and MOM metabolomes determined that over one thousand features were significantly different. In addition, significant changes in the metabolite concentrations were observed in MOM and RM30 samples after four hours of incubation, while the concentration of metabolites in DHM remained constant, indicating that these changes are related to the microbial expansion. In summary, our analyses indicate that the metabolite profiles of DHM are significantly different from that of MOM, and the profile of MOM may be partially restored in DHM through microbial expansion.

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Inadequate Content of Docosahexaenoic Acid (DHA) of Donor Human Milk for Feeding Preterm Infants: A Comparison with Mother’s Own Milk at Different Stages of Lactation

Nutrients ◽

10.3390/nu13041300 ◽

2021 ◽

Vol 13 (4) ◽

pp. 1300

Author(s):

Félix Castillo ◽

Félix-Joel Castillo-Ferrer ◽

Begoña Cordobilla ◽

Joan Carles Domingo

Keyword(s):

Fatty Acids ◽

Docosahexaenoic Acid ◽

Human Milk ◽

Linolenic Acid ◽

Preterm Infants ◽

High Dose ◽

Mature Milk ◽

Donor Human Milk ◽

Milk Banking ◽

Mother’S Own Milk

A cross-sectional single-center study was designed to compare the fatty acids profile, particularly docosahexaenoic acid (DHA) levels, between milk banking samples of donor human milk and mother’s own milk (MOM) for feeding preterm infants born before 32 weeks’ gestation. MOM samples from 118 mothers included colostrum (1–7 days after delivery), transitional milk (9–14 days), and mature milk (15–28 days and ≥29 days). In the n-3 polyunsaturated fatty acids (PUFAs) group, the levels of α-linolenic acid (C18:3 n3) and DHA (C22:6 n3) showed opposite trends, whereas α-linolenic acid was higher in donor human milk as compared with MOM, with increasing levels as stages of lactation progressed, DHA levels were significantly lower in donor human milk than in MOM samples, which, in turn, showed decreasing levels along stages of lactation. DHA levels in donor human milk were 53% lower than in colostrum. Therefore, in preterm infants born before 32 weeks’ gestation, the use of pasteurized donor human milk as exclusive feeding or combined with breastfeeding provides an inadequate supply of DHA. Nursing mothers should increase DHA intake through fish consumption or nutritional supplements with high-dose DHA while breastfeeding. Milk banking fortified with DHA would guarantee adequate DHA levels in donor human milk.

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Inoculation of Mother’s Own Milk Could Personalize Microbiological, Metataxonomic and Peptidomic Profile of Pasteurized Donor Human Milk Used for Feeding Preterm Infants

10.21203/rs.3.rs-439999/v1 ◽

2021 ◽

Author(s):

Domenica Mallardi ◽

Chiara Tabasso ◽

Pasqua Piemontese ◽

Stefano Morandi ◽

Tiziana Silvetti ◽

...

Keyword(s):

Human Milk ◽

Preterm Infants ◽

Bacterial Growth ◽

Bacterial Count ◽

Total Bacterial Count ◽

Milk Samples ◽

Donor Human Milk ◽

Peptidomic Analysis ◽

Mother’S Own Milk

Abstract Background Human milk is a vehicle of bioactive compounds and beneficial bacteria which promote the establishment of a healthy gut microbiome of newborns, especially of preterm infants. Pasteurized donor human milk (PDHM) is the second-best option when preterm mother’s own milk is unavailable. Since pasteurization affect the quality of donor milk, the effect on bacterial growth, human milk microbiota diversity and proteolytic phenomena in PDHM inoculated with different preterm milk samples, was evaluated. Methods Ten preterm milk samples was used to perform inoculation of PDHM at 10% v/v. Microbiological, metataxonomic and peptidomic analysis, on preterm milk samples at the baseline (T0) and PDHM and inoculated milk (IM) samples at T0, after 2h (T1) and 4h (T2) of incubation at 37°C, were conducted. Results IM samples at T2 showed a Total Bacterial Count not significantly different (p > 0.01) compared to preterm milk samples. Lactic acid bacteria (LAB) level was restored in all IM at T2. After inoculation, metataxonomic analysis showed that Proteobacteria remained the predominant phylum while Firmicutes moved from 3% at T1 to 9.4% at T2 in IM samples. Peptidomic profile of IM resembled that of PDHM incubated for the same time in terms of number and type of peptides. Conclusion The study demonstrated that inoculation with fresh mother’s own milk could restore bacterial growth and personalize human milk microbiome in PDHM. This effect could be beneficial thanks to the presence of maternal probiotic bacteria which make PDHM more similar to mother’s own milk.

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A clinical audit of the growth of preterm infants fed predominantly pasteurised donor human milk v. those fed mother’s own milk in the neonatal intensive care unit

British Journal Of Nutrition ◽

10.1017/s0007114519000357 ◽

2019 ◽

Vol 121 (09) ◽

pp. 1018-1025 ◽

Cited By ~ 3

Author(s):

Megan L. Lloyd ◽

Eva Malacova ◽

Ben Hartmann ◽

Karen Simmer

Keyword(s):

Human Milk ◽

Preterm Infants ◽

Gestational Age ◽

Growth Velocity ◽

Neonatal Intensive Care ◽

Clinical Audit ◽

Donor Human Milk ◽

Weight Growth ◽

Significant Difference ◽

Mother’S Own Milk

AbstractPreterm infants whose mothers are unable to produce sufficient breast milk are increasingly being supplemented with pasteurised donor human milk (PDHM) instead of commercial preterm infant formula. Concerns have been raised that this practice can result in reduced growth. This retrospective clinical audit collected data from the medical records of a cohort of preterm infants (≤30 weeks gestational age) receiving either ≥28 d of PDHM (n 53) or ≥28 d of their mother’s own milk (MOM, n 43) with standard fortification supplied to both groups during admission. Weight growth velocity was assessed from regained birth weight to 34+1 weeks’ postmenstrual age (PMA); and weight, length and head circumference were compared at discharge and 12 months (corrected age). At 34+1 weeks’ PMA, the weight growth velocity (g/kg per d) was significantly lower in the PDHM group (15·4 g/kg per d, 95 % CI 14·6, 16·1) compared with the MOM group (16·9 g/kg per d, 95 % CI 16·1, 17·7, P=0·007). However, the increase was still within clinically acceptable limits (>15 g/kg per d) and no significant difference was observed in the weight between the two groups. There was no significant difference in weight between the groups at discharge or at the 12-month corrected gestational age review. Although we demonstrated a significant reduction in the weight growth velocity of preterm infants receiving PDHM at 34 weeks’ PMA, this difference is not present at discharge, suggesting that the growth deficit is reduced by supplementation before discharge.

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Tolerance of preterm formula versus pasteurized donor human milk in very preterm infants: a randomized non-inferiority trial

Italian Journal of Pediatrics ◽

10.1186/s13052-018-0532-7 ◽

2018 ◽

Vol 44 (1) ◽

Cited By ~ 5

Author(s):

Simonetta Costa ◽

Luca Maggio ◽

Giovanni Alighieri ◽

Giovanni Barone ◽

Francesco Cota ◽

...

Keyword(s):

Human Milk ◽

Preterm Infants ◽

Very Preterm Infants ◽

Very Preterm ◽

Donor Human Milk ◽

Preterm Formula

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Stool Microbiota in Infants Receiving Extensively Hydrolyzed Formula, Amino Acid Formula, or Human Milk Through Two Months of Age (FS04-07-19)

Current Developments in Nutrition ◽

10.1093/cdn/nzz048.fs04-07-19 ◽

2019 ◽

Vol 3 (Supplement_1) ◽

Author(s):

Car Reen Kok ◽

Bradford Brabec ◽

Maciej Chichlowski ◽

Cheryl Harris ◽

Nancy Moore ◽

...

Keyword(s):

Amino Acid ◽

Human Milk ◽

Gut Microbiome ◽

Beta Diversity ◽

Alpha Diversity ◽

Rrna Genes ◽

Infant Formulas ◽

Diversity Measures ◽

Mother’S Own Milk ◽

Stool Microbiota

Abstract Objectives Infant feeding practices play a central role in development of gut microbiome and community structure. Our goal was to test the hypothesis that diets with intact or extensively hydrolyzed proteins or free amino acids may differentially affect the intestinal microbiota composition and immune reactivity. Methods This multicenter, double-blind, controlled, parallel-group, pilot study compared stool microbiota outcomes from Baseline (1-7 days of age) up to 60 days of age in healthy term infants. Infants received mother's own milk (assigned to human milk [HM] reference group) (n = 25) or were randomized to receive one of two infant formulas: amino-acid based (AAF; n = 25) or extensively hydrolyzed cow's milk protein (EHF; n = 28). Neither study formula included added Lactobacillus rhamnosus GG. DNA was extracted (Baseline, Day 30, Day 60), 16S rRNA genes were amplified and sequenced (Illumina MiSeq), and exact amplicon sequence variants (ASV) were assigned using the DADA2 model. Alpha (Shannon, Simpson, Chao1) and beta diversity (Bray Curtis distance) and differential abundance in taxa were analyzed. Relative ASV enrichment (Baseline vs Day 60) was visualized using heat maps and taxa abundance was analyzed by DESEq2 in R (ver 3.4.3). Results Complete stool data (all study time points) were available for 49 participants. Baseline alpha diversity measures were similar among groups. The HM group remained stable throughout the study. However, alpha diversity measures by Day 60 were significantly higher for AAF and EHF groups compared to HM. Significant group differences in beta diversity at Day 60 were detected (P < 0.001); AAF and EHF clustered more closely compared to the HM group. Relative Bifidobacterium abundance increased over time and was significantly enriched at Day 60 in the HM group (Figure, A). At Day 60, a significant increase in members of Firmicutes was detected for AAF and EHF groups; a decrease in Enterobacteriaceae (Escherichia) was observed for EHF (Figure, B). Conclusions Distinct patterns of early neonatal microbiome establishment were demonstrated for infants receiving mother's own milk compared to amino acid-based or extensively hydrolyzed protein infant formulas. Providing different sources of dietary protein early in life may impact gut microbiome development. Funding Sources Mead Johnson Pediatric Nutrition Institute. Supporting Tables, Images and/or Graphs

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Impact of Donor Human Milk in the Preterm Very Low Birth Weight Gut Transcriptome Profile by Use of Exfoliated Intestinal Cells

Nutrients ◽

10.3390/nu11112677 ◽

2019 ◽

Vol 11 (11) ◽

pp. 2677 ◽

Cited By ~ 1

Author(s):

Anna Parra-Llorca ◽

María Gormaz ◽

Sheila Lorente-Pozo ◽

Maria Cernada ◽

Ana García-Robles ◽

...

Keyword(s):

Human Milk ◽

Preterm Infants ◽

Very Low Birth Weight ◽

Principal Component ◽

Postnatal Period ◽

Intestinal Cells ◽

Donor Human Milk ◽

Genome Wide ◽

Alpha Gene ◽

Cytosolic Factor

Background: Own mother’s milk (OMM) is the optimal nutrition for preterm infants. However, pasteurized donor human milk (DHM) is a valid alternative. We explored the differences of the transcriptome in exfoliated epithelial intestinal cells (EEIC) of preterm infants receiving full feed with OMM or DHM. Methods: The prospective observational study included preterm infants ≤ 32 weeks’ gestation and/or ≤1500 g birthweight. Total RNA from EEIC were processed for genome-wide expression analysis. Results: Principal component analysis and unsupervised hierarchical clustering analysis revealed two clustered groups corresponding to the OMM and DHM groups that showed differences in the gene expression profile in 1629 transcripts. The OMM group overexpressed lactalbumin alpha gene (LALBA), Cytochrome C oxidase subunit I gene (COX1) and caseins kappa gene (CSN3), beta gene (CSN2) and alpha gene (CSN1S1) and underexpressed Neutrophil Cytosolic Factor 1 gene (NCF1) compared to the DHM group. Conclusions: The transcriptomic analysis of EEIC showed that OMM induced a differential expression of specific genes that may contribute to a more efficient response to a pro-oxidant challenge early in the postnatal period when preterm infants are at a higher risk of oxidative stress. The use of OMM should be strongly promoted in preterm infants.

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