Living bacteria are used in biotechnologies that lead to improvements in health care, agriculture, and energy. Encapsulating bacteria into flexible and modular electrospun polymer fabrics that maintain their viability will further enable their use. This review will first provide a brief overview of electrospinning before examining the impact of electrospinning parameters, such as precursor composition, applied voltage, and environment on the viability of encapsulated bacteria. Currently, the use of nanofiber scaffolds to deliver live probiotics into the gut is the most researched application space; however, several additional applications, including skin probiotics (wound bandages) and menstruation products have also been explored and will be discussed. The use of bacteria-loaded nanofibers as seed coatings that promote plant growth, for the remediation of contaminated wastewaters, and in energy-generating microbial fuel cells are also covered in this review. In summary, electrospinning is an effective method for encapsulating living microorganisms into dry polymer nanofibers. While these living composite scaffolds hold potential for use across many applications, before their use in commercial products can be realized, numerous challenges and further investigations remain.