Exploring the effect of vitamin D and DHA supplementation on the urine metabolome of preterm infants by 1H NMR-based metabolomics

Background and objectives: Vitamin D and docosahexaenoic acid (DHA) insufficiency and deficiency could potentially have a great impact on health outcomes in preterm infant. Due to the importance of early nutrition intervention in this population and given the lack of metabolomic studies concerning the supplementations effect on the metabolome of preterm infants, 44 premature infants were studied, divided in two groups, one receiving only vitamin D (DS) and the second both vitamin D and DHA (D-DHAS) supplementation. Two were the main objectives of the study: 1) to look at changes over time in the urinary metabolic profiles of infants before and over two months of supplementation; 2) to compare the urinary metabolome of the two groups after supplementation. Methods: 1H NMR-based metabolomics approach was used to analyze urine samples obtained from preterm newborns at three different time points: at the time of hospital discharge and before supplementation (T0), 1 month (T1) and 2 months (T2) after the beginning of supplementation. Results: A clear temporal dynamics of the urinary metabolic profiles of preterm infants was highlighted by OPLS analysis. Both groups were characterized by growing levels of betaine, N,N-dimethylglycine, creatinine, creatine and guanidinoacetate and diminishing levels of myo-inositol and hydroxyproline with increasing postmenstrual age (PMA). Additionally, for D-DHAS citrate and dimethylamine increased, while lactate decreased over time. OPLS-DA clearly discriminated the two groups after two months of supplementation. Compared to DS, D-DHAS group was characterized by higher levels of betaine, N,N-dimethylglycine, creatinine and dimethylamine and lower amounts of lactate and myo-inositol. Conclusions: Metabolomic analysis of urine from the neonatal period could be a useful tool to understand metabolic processes linked to early nutrition and supplementation. According to our results, vitamin D supplementation exerts in preterm newborns positive effects evaluated with urinary metabolomics. Moreover, it seems that the supplementation with vitamin D and DHA exerts a higher antioxidant and protective action on newborns, and it could also positively affect the body fat composition

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

Fortification of human milk (HM) for preterm and very low-birth weight (VLBW) infants is a standard practice in most neonatal intensive care units. The optimal fortification strategy and the most suitable protein source for achieving better tolerance and growth rates for fortified infants are still being investigated. In a previous clinical trial, preterm and VLBW infants receiving supplementation of HM with experimental donkey milk-based fortifiers (D-HMF) showed decreased signs of feeding intolerance, including feeding interruptions, bilious gastric residuals and vomiting, with respect to infants receiving bovine milk-based fortifiers (B-HMF). In the present ancillary study, the urinary metabolome of infants fed B-HMF (n = 27) and D-HMF (n = 27) for 21 days was analyzed by1H NMR spectroscopy at the beginning (T0) and at the end (T1) of the observation period. Results showed that most temporal changes in the metabolic responses were common in the two groups, providing indications of postnatal adaptation. The significantly higher excretion of galactose in D-HMF and of carnitine, choline, lysine and leucine in B-HMF at T1 were likely due to different formulations. In conclusion, isocaloric and isoproteic HM fortification may result in different metabolic patterns, as a consequence of the different quality of the nutrients provided by the fortifiers

Clinical impact of human breast milk metabolomics

Metabolomics is a research field concerned with the analysis of metabolome, the complete set of metabolites in a given cell, tissue, or biological sample. Being able to provide a molecular snapshot of biological systems, metabolomics has emerged as a functional methodology in a wide range of research areas such as toxicology, pharmacology, food technology, nutrition, microbial biotechnology, systems biology, and plant biotechnology. In this review, we emphasize the applications of metabolomics in investigating the human breast milk (HBM) metabolome. HBM is the recommended source of nutrition for infants since it contains the optimal balance of nutrients for developing babies, and it provides a range of benefits for growth, immunity, and development. The molecular mechanisms beyond the inter- and intra-variability of HBM that make its composition unique are yet to be well-characterized. Although still in its infancy, the study of HBM metabolome has already proven itself to be of great value in providing insights into this biochemical variability in relation to mother phenotype, diet, disease, and lifestyle. The results of these investigations lay the foundation for further developments useful to identify normal and aberrant biochemical changes as well as to develop strategies to promote healthy infant feeding practice

NMR-based metabolomics analysis of organic and conventionally produced formula milk: preliminary results

Nutrition in early life has important biological effects on immediate and lifetime health. In the light of these considerations, products such as specialized and standard infant formulas substitute for human milk have the potential to influence health outcomes differently depending on their composition. The recent knowledge of the long-term health benefits of breast-feeding has addressed research toward the creation of formulas ever closer to the needs of the infant both in term of nutritional and functional compounds. In this regard, metabolomics has proved to be a promising tool to investigate the metabolic composition of breast milk and the differences compared with formula milk. To the best of our knowledge, no metabolomics studies on the compositional differences between organic and conventionally produced infant milk have been performed so far. To fill this gap, the aim of the present work was to use the 1H NMR-based metabolomics approach to compare the metabolome of organic and conventionally produced formula milk designed for fulfill infants’ nutritional needs from birth to 12 months of age. Methionine content was found to be significantly (p = 0.001) higher in organic milk than in conventional formulas. For the sake of comparison, the metabolome of human milk samples was also analyzed. Although the study presents several limitations, our preliminary results further support the utility of metabolomics in research for infant nutrition

Metabolomics of breast milk: The importance of phenotypes

Breast milk is the gold standard of nutrition for newborns. Its composition is tailored to the nutritional needs of the infant and varies between mothers. In recent years, several bioactive molecules have been discovered in addition to the main nutrients, such as multipotent stem cells, hormones, immunoglobulins, and bacteria. Furthermore, the human milk oligosaccharides (HMOs) seem to exert several important protective biological functions. According to the HMOs’ composition, breast milk can be classified as a secretory or non-secretory phenotype. In our study, we investigated the metabolome of milk collected from 58 mothers that delivered neonates at term, that were appropriate, small or large for gestational age, by performing nuclear magnetic resonance spectroscopy (1 H-NMR). From the data analysis, two groups were distinguished based on their different types of oligosaccharides, and classified according the mother phenotype: secretory and non-secretory. This information is of major importance given the different biological function of the different HMOs, such as immune-modulation and protection against disease. This would allow us to predict whether the neonate would be, for instance, more prone to developing certain diseases, and to tailor her or his nutrition to fit their needs perfectly and pave the way to a personalized nutrition

A Comparison of Mother’s Milk and the Neonatal Urine Metabolome: A Unique Fingerprinting for Different Nutritional Phenotypes

The ability of metabolomics to provide a snapshot of an individual’s metabolic state makes it a very useful technique in neonatology for investigating the complex relationship between nutrition and the state of health of the newborn. Through an 1H-NMR metabolomics analysis, we aimed to investigate the metabolic profile of newborns by analyzing both urine and milk samples in relation to the birth weight of neonates classified as AGA (adequate for the gestational age, n = 51), IUGR (intrauterine growth restriction, n = 14), and LGA (large for gestational age, n = 15). Samples were collected at 7 ± 2 days after delivery. Of these infants, 42 were exclusively breastfed, while 38 received mixed feeding with a variable amount of commercial infant formula (less than 40%) in addition to breast milk. We observed a urinary spectral pattern for oligosaccharides very close to that of the corresponding mother’s milk in the case of exclusively breastfed infants, thus mirroring the maternal phenotype. The absence of this good match between the infant urine and human milk spectra in the case of mixed-fed infants could be reasonably ascribed to the use of a variable amount of commercial infant formulas (under 40%) added to breast milk. Furthermore, our findings did not evidence any significant differences in the spectral profiles in terms of the neonatal customize centile, i.e., AGA (adequate for gestational age), LGA (large for gestational age), or IGUR (intrauterine growth restriction). It is reasonable to assume that maternal human milk oligosaccharide (HMO) production is not or is only minimally influenced by the fetal growth conditions for unknown reasons. This hypothesis may be supported by our metabolomics-based results, confirming once again the importance of this approach in the neonatal field

Impact of Early Postnatal Nutrition on the NMR Urinary Metabolic Profile of Infant

NMR-based metabolomics was used to compare the metabolic urinary profiles of exclusively breast-fed term infants (n = 11) with those of a double-blinded controlled trial with 49 formula-fed term newborns randomized to receive either an infant formula enriched by functional ingredients (n = 24) or a standard formula (n = 25). Anthropometric measurements and urine samples were taken at enrollment (within the first month of life), at around 60 days of life, and at the end of study period (average age of 130 days). The metabolic profiles were examined in relation to time and diet strategy. A common age-dependent modification of the urine metabolome was observed for the three types of nutrition, mainly characterized by similar temporal trends of choline, betaine, myoinositol, taurine, and citrate. Contrariwise, differences in the metabolic profiles were identified according to the type of diet (human versus formula milk), while no significant difference was observed between the two formulas. These modifications are discussed mainly in terms of the different milk compositions. Despite the low number of enrolled infants (n = 60), these findings pointed out the potential of the metabolomics approach for neonatal nutritional science, in particular to provide important contributions to the optimization of formula milk