Iron deficiency is a global health problem, particularly affecting women of reproductive age and young children in developing regions. An estimated 1.62 billion people suffer from anemia, the main cause of which is iron deficiency. Despite various interventions, including supplementation and food fortification, challenges in access and compliance undermine their effectiveness. Biofortification has emerged as a sustainable solution to increase the nutritional value of staple crops and animal products, such as meat and poultry, which are important sources of bioavailable heme iron. This chapter explores methodologies of iron biofortification in livestock, highlighting effective management practices, agronomic strategies, and genetic approaches. Key techniques include a selection of iron-rich species, use of iron-rich feed, and soil amendment to improve iron content in forages. Additionally, genetic modification and selective breeding are being explored as innovative approaches to enhance iron absorption and retention characteristics in livestock. Laboratory protocols for measuring iron levels in animal tissues and blood samples are also described, ensuring accurate evaluation of biofortification efforts. By combining these strategies, this chapter offers insight into promising pathways to address iron deficiency and improve public health outcomes, particularly in vulnerable populations.