Saidatul Akasha Sufian, Syed Muhammad Omer Khalid, Mohamad Syahmi Iskandar Sahran,

Seok Fang Oon and Hajar Fauzan Ahmad

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MOLECULAR IDENTIFICATION OF LACTIC ACID BACTERIA ISOLATED FROM STINGLESS BEE HONEY IN PAHANG, MALAYSIA 

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Abstract 

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Stingless bee honey produced by Heterotrigona itama is recognized as a natural habitat rich in diverse microorganisms, including lactic acid bacteria (LAB) with potential probiotic properties. However, studies exploring the microbial diversity and molecular identification of bacteria in local stingless bee honey from Malaysia remain limited. This study aimed to isolate and identify potentially probiotic bacterial isolates from raw stingless bee honey collected from Rimba Lestari, Universiti Malaysia Pahang Al-Sultan Abdullah (UMPSA), Pekan. The samples were aseptically inoculated onto MRS agar and incubated aerobically at 37°C for 48–72 hours. A total of four bacterial isolates were successfully obtained and identified as Gram-positive, rod-shaped bacteria through Gram staining. Partial 16S rRNA gene sequencing using the Sanger method revealed that two isolates (UMPSA1 and UMPSA2) showed high similarity (>99%) to uncultured Bacillus species, while the other two isolates (UMPSA3 and UMPSA4) shared 100% identity with uncultured bacteria from the order Bacillales. Although no isolates from LAB genera such as Lactobacillus or Fructobacillus were recovered, the presence of Bacillus is notable, as several strains from this genus have been reported to possess probiotic traits, such as enzyme production and resilience in the gastrointestinal environment. One key limitation of this study is the use of short gene sequences, which restricted the accuracy of species-level identification. Future research is recommended to apply Whole Genome Sequencing (WGS), alongside the use of lactose- or fructose-enriched selective media and incubation under anaerobic or microaerophilic conditions, to enhance the isolation and characterization of novel LAB strains with probiotic potential from stingless bee honey.

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