In Myanmar, 60% of the population consists of mothers and children, and they are the groups most vulnerable to anemia. The objectives of this study are to determine (1) the anemia prevalence among lactating women and (2) the risk factors associated with anemia. Convenience sampling was used to select three villages in two different regions (Kachin and Shan) in Myanmar. Hemoglobin and anthropometric indicators were measured for 733 lactating women. Logistic regression analyses were used to determine factors associated with anemia. The anemia prevalence rate was 60.3% in lactating women, with 20.3% of lactating women having severe anemia. Factors of malnutrition (P = 0.026), self-reported symptoms of night blindness or poor dark adaptation (P < 0.001), lack of primary education experience (P < 0.001), low family annual capita income (< 800 MMK; P < 0.001), drinking spring or river water (P < 0.001), and drinking unboiled water (P = 0.016) were associated with anemia. To promote health in lactating women, a comprehensive intervention is needed in these regions.