Investigating how dietary nutrients influence the structure and action of the gut flora is a topic of significant importance in nutrition and health. The gut microbiota is accountable for the production of various bioactive chemicals, among which are B vitamins. These vitamins play crucial physiological functions in the host organism by promoting symbiotic species' well-being and inhibiting competing species' proliferation. B vitamins have a crucial role in the well-being of both the host and certain gut bacteria. Nutritious B vitamins are mostly absorbed in the small colon; however, any surplus B vitamins that cannot be absorbed in this region are sent to the distal gut. Moreover, the distal gut microbiota oversees the manufacture of B vitamins that are present in the distal colon and are known to serve various crucial roles within the body. These roles encompass acting as essential nutrients for both the host and their microbiota, regulating immune cell activity, influencing the effectiveness of certain drugs, promoting the survival and fitness of specific bacteria, inhibiting the colonization of pathogenic bacteria, and modulating colitis. In this chapter, we will examine the significance of individual B vitamins in the distal intestine, focusing on their interactions with bacteria and mechanisms. The exploration of expanding analytical tools to enhance the comprehension of the gut's involvement in B vitamins is being investigated. The host's metabolism is greatly affected by deficits in these vitamins due to their widespread functions. In recent times, there has been emerging research on the unique B vitamins functions in maintaining the balance of gut Bacillus science and promoting intestinal well-being.