Medicon and C-TRIC

“Expressed Human Breast Milk Microbiota Study (EHBMMS)”

Expressed Human Breast Milk Microbiota Study (EHBMMS) is a collaboration between three organisations – C-TRIC, the Human Milk Bank at SWAH and Medicon Ireland.

The Western Trust Milk Bank opened in August 2000. Since then it has helped thousands of neonatal babies all over Ireland. In 2021 the milk bank processed a total donation of 2,142 litres of donor expressed milk and have provided approximately 1,427 litres of breast milk to 27 neonatal units in hospitals across Ireland, helping 558 babies. The bank also helps babies with major gut or heart problems by providing milk to their home.  Through the generosity of its donors, the milk bank provides very special help for the nation’s most vulnerable babies.

Medicon Ireland, Established in January 2007 by Managing Director, Iggy McGowan, are one of Ireland’s fastest growing companies within the Diagnostic and Wholesale Pharmaceutical Sectors. Specialising in laboratory diagnostic equipment, point of care systems, blood glucose meters and pharmaceuticals.

The human microbiome and its relevance to health and development is becoming increasingly recognised in the scientific community.  In order to further the understanding of how the microbiome relates to health status, it is imperative that researchers and healthcare providers have access to a reliable and accurate diagnostic process. Studies consistently find the beneficial effects of breastfeeding on overall maternal and infant health with breast-fed infants have a significantly lower incidence of respiratory infection, diarrhoea, and dehydration, as well as fewer signs and symptoms of atopic disease (Binns et al., 2016; LA et al., 2003; P et al., 2019). These health advantages underscore the importance of maintaining the breastfeeding relationship for mother and infant for as long as possible. Human milk is fundamental for the correct development of newborns, as it is a source not only of vitamins and nutrients, but also of commensal bacteria (Collado et al., 2009). The microbiota associated with human breast milk contributes to create the “initial” intestinal microbiota of infants, also having a pivotal role in modulating and influencing the new-borns’ immune system. Bacteria located in both colostrum and mature milk can stimulate the anti-inflammatory response, by stimulating the production of specific cytokines, reducing the risk of developing a broad range of inflammatory diseases and preventing the expression of immune-mediated pathologies, such as asthma and atopic dermatitis (Binns et al., 2016; LA et al., 2003). Human breast milk is recognized as the best source of nutrients for healthy growth and development of infants because it contains bioactive components, including oligosaccharides which feed and stimulate the growth and activity of good bacteria in the gastrointestinal system of the infant (Pacheco et al., 2015). Bacteria which pass from the mother to infant via breast milk has a large effect on the makeup of the gut microbiome in the baby. Therefore, testing the mother’s breast milk microbiota would unearth any deficiencies in its bacterial makeup which then could be addressed by a healthcare professional/clinician.

Our Microbiome Analysis combined sample collection and processing, NextGen sequencing and bioinformatics analysis to provide unprecedented details of the composition of microbial species within human breast milk. Microbiome taxonomic profiling (MTP) is a method to determine the bacterial composition of a sample and are commonly performed by analysing the prokaryotic 16S ribosomal RNA gene (16S rRNA), which is approximately 1,500 bp long and contains nine variable regions interspersed between conserved regions. Variable regions of 16S rRNA are frequently used in phylogenetic classifications such as genus or species in diverse microbial populations. This protocol describes a method for preparing samples for targeted sequencing of the variable V3 and V4 regions of the 16S rRNA gene.

The project to date has involved the development and optimisation of a 16s ribosomal RNA sequencing protocol to identify and quantity bacterial species within human breast milk samples. In addition, corresponding immunological/nutritional assays have been developed to correlate bacteria which appear to be beneficial for newborns immune system and development. The multi-year project plans to create a diagnostic test for mothers’ breast milk to have access to a reliable, robust and accurate breast milk analysis service to analyse the nutritional and immunological qualities of their breastmilk, this will allow mothers’ and clinicians to make informed decisions for breastfeeding and newborn nutrition.

Important ethical considerations have been given to reduce sample size, simplify sample storage and transportation, accelerate existing protocols, enhance the quality of results and make the process affordable. At all stages, sensitivities must be shown to the mother and GDPR guidelines must be followed.