Probiotics play a pivotal role in health maintenance and prevention of many disorders. They are microbial feed supplements that confer the health effect on the host. Probiotics protects the body itself from infection, especially gastrointestinal tract. Acidophilus is a probiotic microorganisms that have a beneficial effect on the host. Lactobacilli acidophilus are "friendly" bacteria that normally live in our digestive, urinary and genital systems without causing disease. The therapeutic benefits have been investigated in women having vaginal and urinary tract infections. In this review, we studied about previous 15 years articles in which the isolation of Lactobacilli bacteria from different sources was mentioned. It has been found that they may be isolated from fruits and vegetables, human stool culture, natural antimicrobial agent, cheese, kefir grains, dairy and non-dairy products, fermented and raw milk, feces of breast fed infants, lactating milk, sheep, buffalo and cow milk, yogurt, beverages, poultry sources, animal rumen contents, pengging duck’s caecum, chicken intestine and fecal samples, chicken feed, enzymes, fermented rice, curd, meat and yeast extracts, glucose and sucrose, human gut, human colonial epithelial cells, human and animal vagina and mouth extraction, diapers of human babies, pineapples wastes, industrial sausages, ice-cream, small intestines of piglets, corn slurry, crop and intestinal ducks. Hence the paper reviews the current scenario of isolation of Lactobacilli from different sources, their proposed mechanisms and health benefits for human beings along with their future perspectives.