Probiotics are considered effective microbial dietary supplements that provide beneficial effects to consumers, usually by restoring or improving gut microflora. Goat milk is one of the rich sources of probiotics as well as nutrients. Therefore, the primary aim of this research was to isolate and evaluate the potential of novel indigenous probiotic strains present in goat milk. Six different raw goat milk samples were collected from different areas of Multan, Pakistan. For bacterial characterization, samples were cultured and isolated on MRS agar plates for different morphological and biochemical tests. The probiotic potential of the six isolates, all of which were gram positive (G1, G2, G3, G4, G5, and G6) and five of which were catalase negative (all except G1), were assessed via a milk coagulation assay and antimicrobial activity, pH tolerance, phenol tolerance, and sodium chloride (NaCl) tolerance tests, which revealed that all the isolates coagulated in milk and showed protease and lipase activity, except G3. All six isolates showed tolerance against 0.2% phenol and 2–4% NaCl and were able to survive in both alkaline and acidic conditions. Only five isolates showed antimicrobial activity against indicator strain Aspergillus niger strain STA9, validating their probiotic nature. The most potent bile-tolerant and bacteriocin-producing isolate, G1, also showed γ-hemolytic activity and resistance to penicillin but showed susceptibility to other antibiotics. The lactic acid-producing (0.60% titratable acidity) G1 isolate was identified as a novel strain of Mammaliicoccus sciuri based on 16S rDNA sequencing. The above findings suggest that the potent M. sciuri GMN01 strain can serve as a potential probiotic strain. A potent probiotic strain isolated from raw goat milk could be utilized as a dietary supplement, and goat milk could become an alternative to other sources of milk, particularly cow milk. However, safety aspects of this strain require further investigation because the present safety tests are insufficient to conclude that the GMN01 isolate is safe.
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