Undergraduate Research Journal for Biomolecular Sciences and Biotechnology

Pemencilan dan Pengenalpastian Bakteria Daripada Madu Kelulut (Trigona sp.)

Lia Amira Said1, Muhammad Azim Izzani2, Nik Marzuki Sidik 3 & Fareed Sairi1* 1Mikrobiologi, Fakulti Sains dan Teknologi, Universiti Kebangsaan Malaysia, 43600 Bangi,

Selangor, Malaysia 2Universiti Sains Malaysia, 11800 Gelugor, Penang, Malaysia

3Fakulti Industri Asas Tani, Universiti Malaysia Kelantan, Kampus Jeli, Beg Berkunci 100, 17600 Jeli, Kelantan,Malaysia

*Mel-e pengarang rujukan: fareed@ukm.edu.my Abstrak: Madu merupakan makanan yang dihasilkan oleh lebah daripada nectar tumbuhan. Selain daripada sumber yang kaya dengan pelbagai nutrien, madu juga menjadi takungan mikroorganisma yang berasal daripada persekitaran. Kajian lepas telah mengenalpasti kehadiran bakteria patogen yang berbahaya dalam madu . Walau bagaimanapun, pengetahuan tentang bakteria dalam madu kelulut pada masa ini masih sangat sedikit. Justeru, kajian ini dijalankan untuk memencilkan dan mengenalpasti bakteria daripada madu kelulut. Sebanyak 19 bakteria berjaya dipencilkan daripada madu kelulut. Kesemua bakteria yang telah dipencilkan telah dicerap secara makroskopik dan mikroskopik. Genom DNA daripada 14 pencilan telah berjaya diekstrak daripada kalangan 19 pencilan bakteria. Tindak balas rantai polimerase (PCR) dan penjujukan telah berjaya dilakukan terhadap lapan pencilan. Produk PCR yang telah dijujuk dan dipadankan menunjukkan kelapan-lapan pencilan bakteria tersebut adalah Bacillus thuringiensis (GTLi 1/2), Bacillus aryabhattai (HTLi1/3), Bacillus aerius (GTLi 1/7 dan GTLi 2/14), Bacillus stratosphericus (GTLi 1/9), Bacillus altitudinis (GTLi 1/13), Bacillus megaterium (GTLi 2/15) dan Bacillus amyloliquefaciens (HTLi 1/19). Pokok filogeni yang telah dibina daripada jujukan pencilan bersama jujukan rujukan daripada GenBank mengesahkan bahawa kesemua bakteria pencilan adalah tergolong dalam divisi Firmicutes dan famili Bacilliceae.

Kata kunci: madu kelulut; Bacillus; bakteria

Abstract: Honey is a food produced by bees rather from plant nectar. Apart from a rich source of nutrients, honey is also a reservoir for microorganisms that come from the environment. Past studies had identified the presence of pathogenic bacteria that are harmful in honey. However, the knowledge of bacteria in today's stingless bee honey is still very little. Hence, this study is conducted to isolate and identify bacteria from stingless bee honey. A total of 19 bacteria was isolated from stingless bee honey. All isolated bacteria were observed macroscopically and microscopically. DNA genomes of 14 isolates were successfully extracted out of 19 bacterial strains. Polymerase chain reaction (PCR) and sequencing were successfully performed on eight strains. PCR products that have been sequenced and matched show that the eight bacterial strains are Bacillus thuringiensis (GTLi 1/2), Bacillus aryabhattai (HTLi1 / 3), Bacillus aerius (GTLi 1/7 and GTLi 2/14), Bacillus stratosphericus (GTLi 1/9), Bacillus altitudinis (GTLi 1/13), Bacillus megaterium (GTLi 2/15) and Bacillus amyloliquefaciens (HTLi 1/19). The phylogenetic tree was constructed from sequences of isolates with the reference sequence of GenBank confirmed that all bacterial isolates were classified in the division Firmicutes and family Bacilliceae.

Keywords: stingless bee honey; Bacillus ; bacteria