Microbial Profiling of Ethnic Fermented Beverages of Himachal Pradesh Using Phenotypic and Molecular Approaches

Abstract

Traditional fermented beverages of the Himalayan region represent an important repository of indigenous knowledge, microbial diversity, and functional nutrition. The present study investigates the bacterial diversity associated with selected ethnic fermented beverages of Upper Himachal Pradesh, India, with emphasis on their molecular identification and potential health relevance. Samples of Angoori, Chaas, Sattu, Chol, and Brandy were collected from different districts and subjected to microbial isolation using serial dilution and spread plate techniques on skim milk agar. Selected bacterial isolates were characterized based on colony morphology, Gram staining, and standard biochemical assays, including catalase, oxidase, methyl red, Voges–Proskauer, and hydrogen sulphide production. Isolates exhibiting desirable biochemical traits and negative H₂S production were further analyzed through 16S rRNA gene amplification and sequencing. Phylogenetic analysis revealed that the dominant isolates belonged to the genus Bacillus, including Bacillus anthracis, Bacillus arachidis, Bacillus velezensis, Bacillus amyloliquefaciens, and closely related Bacillus spp. The findings highlight the microbial richness of these traditional beverages and underline the limitations of 16S rRNA sequencing in resolving closely related species. Overall, this study provides scientific validation of the microbial composition of Himalayan fermented beverages and supports their potential as functional foods with probiotic and health-promoting attributes, while emphasizing the need to conserve traditional fermentation practices and associated microbial resources.