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The concept of probiotics evolved more than a century ago, linking longevity with the consumption of certain Lactobacillus strains present in yogurt. Ever since, the known health benefits of probiotics have been mounting with the advancements in scientific technologies. Benefits of probiotics consumption extend far beyond gut health, to improving conditions of allergies, mental health such as depression and anxiety, wound healing, menopause, bacterial infections, antibiotic resistance, etc. Scientific studies have established that the efficacy of probiotics is strain-specific. These strains are made available to consumers for use in different forms, viz., functional foods, dietary supplements, and probiotic drugs for human consumption. In order to monitor and regulate the use of these probiotics for human/animal/drug consumption, there are existing guidelines and a regulatory framework. As per these guidelines, for a microorganism to be claimed as a probiotic, certain criteria need to be fulfilled, such as strain characterization, safety evaluation, and efficacy. This chapter provides an overview of probiotics and the details of safety assessment requirements for the usage of probiotics in food. It also covers the details of the regulatory framework in India for the use of probiotics in food, the labeling and health claim requirements as notified by the Food Safety and Standards Authority of India (FSSAI), and provides a short summary of guidance for animal feed and pharma use.