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

Abstract

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.

A Pilot Study on Donor Human Milk Microbiota: A Comparison with Preterm Human Milk Microbiota and the Effect of Pasteurization

“…Regarding Pseudomonas, our result is consistent with studies showing a higher relative abundance of Pseudomonadaceae in the HM of mothers of term infants who reported using a breast pump compared with mothers who did not [32]. On the other hand, our finding on alpha diversity is in contrast with previous reports, showing similar values between DHM and PT-MOM samples [34,41]. It must be said that the lactation stage has also been shown to have an impact on the HM microbiota [42].…”

“…With regard to pasteurization, evidence has been accumulating over the last years on the effects of HoP on milk nutrients and bioactive components, but only limited data on the HM microbiota have been reported [20,34,41]. Since it is an effective means of removing any potentially pathogenic bacteria, HoP is thought to most likely kill much of the HM microbiota.…”

Lactobacillus johnsonii N6.2 and Blueberry Phytophenols Affect Lipidome and Gut Microbiota Composition of Rats Under High-Fat Diet

“…DNA was extracted from stool samples collected from ileum and colon from all animals from all groups using QIAamp ® PowerFecal ® Pro DNA kit (QIAGEN), and preserved at −80°C. The following modification was applied to the extraction protocol: 100 μL of protease (20 mg/mL) from Streptomyces griseus (Sigma–Aldrich, Steinheim, Germany) was added to improve the lysis step ( 35 ). The mixture was incubated at 37°C for 15 min, then the samples were processed according to the kit protocol.…”

Nitrogen metabolism of the infant gut microbiota: a new focus on human milk prebiotics

“…There is more nitrogen in HM, besides the non-protein part there is of course protein. Lactoferrin (LF) is among the most detected proteins in the early life gut, indicating its availability to the microbiota and showing the potential for this (±700 AAs) protein to have a beneficial effect in lowering pathogen colonization [184,185]. Products of the bacterial colonists of the infant gut could be crucial vitamins.…”