IntroductionDietary sources have an abundance of bacteria, mainly lactic acid bacteria (LABs), which have long been regarded as probiotics in humans and animals. Lactic acid bacteria (LAB) have been used as probiotic agents due to their ability to produce a variety of beneficial compounds for cultivars and their status as safe microorganisms.MethodsIn this current study, the lactic acid bacteria (LAB) were isolated from several dietary sources such as curd, pickle, milk, and wheat dough. The principal focus of this study was to determine the survivability of these microorganisms in the gastrointestinal tract and to use promising strains to create probiotic drinks with numerous health benefits. The isolates were identified using a combination of morphological, biochemical, molecular and sugar fermentation patterns, like phenotypic characteristics, sugar fermentation, MR-VP reaction, catalase test, urease test, oxidase test, H2S production, NH3production synthesis from arginine, citrate utilization, indole test, and 16s rRNA sequencing.ResultsTwo (CM1 and OS1) of the 60 isolates obtained showed the best probiotic results and were identified as Lactobacillus acidophilus CM1 andLactobacillus delbrueckiiOS1. These organism sequences were submitted to Gen bank with accession numbers OP811266.1 and OP824643.1, respectively. The acid tolerance test results indicated that most strains could survive significantly in an acidic environment with pH levels of 2 and 3. Similarly, the salt tolerance test results showed that bothLactobacillus acidophilusCM1 andLactobacillus delbrueckiiOS1 could survive at 4 and 6% NaCl levels significantly. The isolates also showed their ability to ferment sugars such as lactose xylose, glucose, sucrose, and fructose.DiscussionIn conclusion, the study showed that the bacteria isolated from different food sources were indeed probiotic lactic acid bacteria and had probiotic properties. These isolates hold potential for future research in the formulation of millet-based probiotic beverages. However, further studies are required to confirm their effectiveness and safety in improving human health. This research provides a foundation for developing functional foods and drinks that can positively affect human health by incorporating probiotic microorganism.