Computational and Microbial Analysis of Metagenome of Commercially Available Cookies Revealed the Presence of Numerous Bacteria

Because of their high nutritional content and extended shelf life, biscuits are eaten all over the world. Because they come in so many shapes, fillings, colors, and topping combinations, biscuits are loved by people of all ages. We have decided to use the computational metagenomic method to investigate commercially available cookies in this study. We selected four commercially available cookies: Cadbury Oreo, Britannia Milk Bikis, Britannia Marie Gold, and Britannia Good Day. Bacteria were isolated from these four cookie brands using the streak plate method. Assays related to microbiology and biochemistry were conducted, including simple staining, gram staining, catalase, methyl red, Vogus Proskauer, TSI, urease, indole, citrate utilization, starch hydrolysis, and antibiotic disc test. The isolates of bacteria were found to be rod-shaped and Gram-positive. The computational metagenomic method revealed that the majority of the trimmed taxa in the sample belonged to the Kingdom Bacteria (73.61%), while 26.38% of the taxa did not have a clear classification (No hit). Within the Bacteria Kingdom, the predominant phylum was Tenericutes (71%), with Cyanobacteria, Proteobacteria, and Firmicutes constituting smaller percentages (1.84%, 0.57%, and 0.13%, respectively). At the class level, Mollicutes represented the highest proportion (71%), followed by Gammaproteobacteria, Alphaproteobacteria, and Betaproteobacteria with smaller percentages (0.34%, 0.11%, and 0.11%, respectively). The other classes had even lower percentages, with Cytophagia, Sphingobacteriia and Bacilli 0.02% each. Moving on to the order level, Acholeplasmatales accounted for the majority (71%), while the other orders had significantly lower percentages, like 0.20% (Cytophagales). Within the families, Acholeplasmataceae made up the largest portion (71%), and the other families had relatively minor representation, ranging from 0.02% (Cytophagaceae) to 0.10% (Paenibacillaceae). At the species level, the dominant taxon was Sugarcane phytoplasma (71%), while the other identified species had much smaller proportions, ranging from 1.84% (Trichodesmium sp) to 0.01% (Hymenobacter metalli and Staphylococcus syringae). Approximately 26.38% of the taxa remained unclassified or did not match any known classification (no hit). Thus, this computational metagenomic study highlights the potential threat by commercially available cookies.