Milk mineral composition is strongly associated with the human milk microbiome

Lilian Lopez Leyva¹Emmanuel Gonzalez^2,3Corinne Maurice^4,5Kristine G Koski^1*

• ¹School of Dietetics and Human Nutrition, Faculty of Agricultural and Environmental Sciences, McGill University, Montreal, Quebec, Canada
• ²McGill Interdisciplinary Initiative in Infection and Immunity (MI4), McGill University, Montreal, QC, Canada
• ³Canadian Centre for Computational Genomics (C3G), Montreal, QC, Canada
• ⁴Microbiology & Immunology Department, McGill University, Montreal, QC, Canada
• ⁵McGill Centre for Microbiome Research, McGill University, Montreal, QC, Canada

Introduction: Associations between maternal mineral intake, human milk mineral concentrations, and their interactions with the milk microbiota remain understudied, especially in low-and middle-income countries. To understand potential interactions and gain insight into milk composition dynamics, we explored associations of milk mineral concentrations with maternal mineral intakes and the human milk microbiome in an indigenous Guatemalan community.Methods: In this cross-sectional study, milk samples were collected from 77 Mam-Mayan mothers and classified into early and established lactation. Concentrations of 9 milk minerals were analyzed, and maternal dietary intake was obtained from two 24-hour recalls. Microbiome diversity was assessed by 16S rRNA gene sequencing (V5-V6 region). DESeq2 was used for differential abundance analysis.PCA and Spearman's rank correlation explored relationships among milk minerals, maternal mineral intake, and differentially abundant microbial taxa; results with FDR-adjusted p-values < 0.1 were retained.Results: Our multifactorial analysis revealed strong associations between milk minerals and the milk microbiome and weak associations with maternal intake. Several maternal intakes (Ca, Se, K, Fe, Mn) and milk mineral concentrations (Ca, Se, K, Mg, Na) were below reference values. In early lactation, milk Fe, Mn, Se, and Cu correlated with differentially abundant taxa, while in established lactation, Fe, Mn, Se, Ca, and Na were correlated. Fe and Mn accounted for 64% of bacterial associations in early lactation and 75% in established lactation. These minerals were correlated with Pseudomonadota (early), Actinomycetota (established), and Bacillota (both), but all species were unique to each stage.Conclusion: Our findings reveal a complex interplay between milk minerals and the microbiome. Iron, manganese, and selenium were consistently associated with milk bacteria across lactation stages. These correlations may reflect microbial responses to mineral availability. Further longitudinal studies with larger samples are needed to clarify how this interaction influences mineral bioavailability and infant growth.