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Human milk oligosaccharides (HMO) are complex carbohydrates that are highly abundant in human milk, but not in infant formula. HMO composition varies between mothers and over the course of lactation. The majority of HMO is indigestible and reaches the distal small intestine and colon intact at high concentrations. Accumulating evidence suggests that HMO benefit the breastfed infant through multiple different mechanisms reviewed in this chapter. HMO have prebiotic effects and shape the microflora composition in the infant's intestine by serving as metabolic substrate for specific, potentially beneficial bacteria. HMO are antiadhesive antimicrobials and serve as soluble decoy receptors that block pathogen attachment to epithelial surfaces to lower the risk for viral, bacterial and protozoan parasite infections; not only in the infant's intestine, but also in the infant's upper respiratory tract and urinary tract and potentially even in the mother's mammary gland. HMO may directly affect epithelial cells, modulate gene expression and alter cell surface glycan expression as well as cell proliferation, differentiation and apoptosis. HMO may modulate the infant's immune system and alter lymphocyte cytokine production, stimulate macrophages and reduce leukocyte extravasation and activation. In animal models, HMO prevent necrotizing enterocolitis, one of the most common and often fatal intestinal disorders in preterm infants. HMO may also provide sialic acid as a potentially essential nutrient for brain development during early postnatal stages. Most of the data related to the beneficial effects of HMO has been generated in in vitro or ex vivo models. Additional research with controlled human intervention studies is required to verify that the postulated effects translate to benefits for the human neonate.