BackgroundVitamin B12 is essential for deoxyribonucleic acid synthesis and genome stability. A deficiency of vitamin B12 is associated with telomere shortening, genomic aging, and increased risk of chronic disease and mortality. ObjectivesThe study aims to determine the effect of vitamin B12 supplementation on leukocyte telomere length (LTL) in infants at risk of vitamin B12 deficiency. MethodsThe study was a predefined secondary analysis of a randomized controlled trial enrolling 600 Nepalese infants aged 6 –11 mo, who were supplemented with 2 μg (2–3 recommended daily allowances) vitamin B12 or placebo daily for 1 y. At the end of the study, LTL was measured in 497 participants. Mean LTL was compared between the treatment arms in the full sample and predefined subgroups based on markers of vitamin B12 status, hemoglobin, sex, and growth indices. ResultsLTL at end-study did not differ between the vitamin B12 and placebo arm with a standardized mean difference (95% confidence interval) of 0.04 (–0.14, 0.21). There was no effect of vitamin B12 on LTL in any of the subgroups. ConclusionsProviding daily vitamin B12 for 1 y during infancy in a population at risk of vitamin B12 deficiency does not affect LTL.This trial was registered at clinicaltrials.gov as NCT02272842.