Unveiling the Microbial Symphony of Amasi: A Targeted Metagenomic 16S rRNA, ITS, and Metabolites Insights Using Bovine and Caprine Milk

Amasi, a traditional fermented milk produced in Southern Africa, is associated with several health benefits, such as probiotic activities, immune system modulation, and pharmacological (antimicrobial, antitumor and antioxidant) potential. This study investigated the microbial diversity in Amasi (produced from cow’s and goat’s milk) through targeted metagenomic bacterial 16S rRNA and fungal ITS sequencing, the metabolic functional prediction of Amasi samples using the Phylogenetic Investigation of Communities by Reconstruction of Unobserved States (PICRUSt) and profiled amino acids constituents using Liquid Chromatographic-Mass Spectrophotometry (LC-MS). The results obtained revealed Firmicutes, Bacteroidetes, and Proteobacteria as the most prevalent bacterial phyla, with Lactococcus and Lactobacillus being the most abundant genera. On the other hand, Ascomycota, Basidiomycota, and Mucoromycota were the main fungal phyla, while Aspergillus, Kazachstania, and Debaryomyces spp. dominated the fungal genera. Also, Pseudomonas spp., Bacillus spp., Clostridium spp., Cronobacter spp., Alternaria spp., Diaporthe spp., and Penicillium spp. were the probable pathogenic bacteria and fungi genera found, respectively. Atopobium, Synechococcus, and Parabacteroides were found less often as rare genera. It was found that the amino acid and drug metabolism pathway prediction values in Amasi samples were significantly higher (p < 0.05) than in raw cow and goat milk, according to the inferred analysis (PICRUSt). The amino acid validation revealed glutamine and asparagine values as the most significant (p < 0.05) for Amasi cow milk (ACM) and Amasi goat milk (AGM), respectively. Comparatively, ACM showed more microbial diversity than AGM, though there were relative similarities in their microbiome composition. PICRUSt analysis revealed significant metabolites in the two Amasi samples. Overall, data from this study showed heterogeneity in microbial diversity, abundance distributions, metabolites, and amino acid balance between raw cow/goat milk and Amasi samples.