Streptococcus pneumoniae is one of the main bacterial causes of pneumonia and is associated with high morbidity and mortality. Fermentation of functional food ingredients containing ginger, lemon, and kelulut honey has the potential to provide LAB with antibacterial activity; however, the characteristics of LAB and their ability to inhibit S. pneumoniae from this fermentation have not been widely investigated. To identify the types of LAB from a fermented beverage made from a mixture of ginger, lemon, and kelulut honey and to determine the antibacterial activity of the isolated LAB against S. pneumoniae. This laboratory experimental study used spontaneous fermentation of a mixture of ginger, lemon, and kelulut honey at fermentation days 0, 1, and 2. LAB isolates were obtained through serial dilution and plating on MRSA medium, then characterized based on colony morphology, Gram staining, and biochemical tests. Antibacterial activity against S. pneumoniae was evaluated using the disk diffusion method on Mueller Hinton Agar. The highest LAB colony count was obtained on day 1 of fermentation, whereas day 0 showed a lower colony count and day 2 showed a subsequent decrease. The isolates consisted of Gram‑positive, non‑spore‑forming, non‑motile cocci on days 0 and 2, phenotypically consistent with the genus Pediococcus, and Gram‑positive, non‑spore‑forming, non‑motile rods on day 1, consistent with the genus Lactobacillus. Disk diffusion testing showed that LAB isolates from days 0, 1, and 2 did not produce inhibition zones against S. pneumoniae. Fermentation of ginger, lemon, and kelulut honey yields LAB candidates phenotypically presumed to belong to the genera Pediococcus and Lactobacillus, with a peak colony count on day 1, however, under the fermentation conditions and antibacterial test method used, antibacterial activity against S. pneumoniae was not detected. Keywords: antibacterial activity, lactic acid bacteria, ginger, lemon, kelulut honey, S. pneumoniae.
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