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Vitamin B12 plays an important role in the mechanisms which are responsible for myelinization in the central nervous system. It can particularly lead to hematological and neuropsychiatric symptoms when serum levels fall due to insufficient intake with diet or absorption problems. The purpose of this study was to show the cognitive effects in vitamin B12 deficiency cases that have not reached clinical symptom level using neuropsychological tests, and to show possible cerebral neuronal damage using diffusion tensor imaging (DTI) method. A total of 62 asymptomatic vitamin B12 deficiency patients and 40 healthy subjects were included in the study and both groups were subjected to Standardized Mini-Mental State Examination, Montreal Cognitive Assessment Test, Rey Auditory Verbal Learning Test, forward and backward digit span (WMS-R forward and backward), Visual Reproduction Subtest (WMS-III), Category Fluency Test, Trail Making (Trail A-B) (21) and Similarities (BENZ) tests. DTI examinations were performed on both groups. Patient group was determined to get lower scores in all neuropsychological tests compared to control group. In DTI examination, a significant decrease in FA values of bilateral hippocampus and a prominent increase in apparent diffusion coefficient (ADC) values were determined in the patient group compared to control group. In this study, it was determined that there was microstructural damage in the brain in the presence of vitamin B12 deficiency even in the asymptomatic period, and the patients revealed cognitive decline. In accordance with this result, early treatment of the easily diagnosed and treated vitamin B12 deficiency may prevent possible irreversible damage in the future.