Abstract: Probiotics are defined as “microbial food supplements” with favorable effects on the consumers. The selection of probiotic microorganisms depends on a number of aspects including safety, functional and technological characteristics. Increased incidence of microbial resistance to antibiotics has triggered renewed efforts to develop probiotics in the treatment of gastric aliments. This review outlines the uses of probiotics and their novel applications and developments for healthcare. The biomedical applications of probiotics in the prophylaxis and treatment of different disease conditions such as diarrhea, constipation, hypercholesterolaemia, vaginitis, necrotizing enterocolitis, sepsis, colon cancer, bladder cancer, breast cancer and liver cancer are discussed. Furthermore, stimulation immune system, weight gain improvement in premature infants, reduction of incidence of respiratory tract infections and prevention of intestinal infections are also highlighted and current challenges are described.