Identifikasi Filogenetik Isolat Bakteri Dari Tambak Garam Osowilangun Berdasarkan Gen 16S rRNA

Risqullah, Muhammad Alvan Thariq (2026) Identifikasi Filogenetik Isolat Bakteri Dari Tambak Garam Osowilangun Berdasarkan Gen 16S rRNA. Other thesis, Institut Teknologi Sepuluh Nopember.

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Abstract

Lingkungan hipersalin seperti tambak garam Osowilangun di Surabaya merupakan ekosistem ekstrem yang berpotensi menjadi habitat mikroorganisme halofilik, termasuk bakteri dengan kemampuan adaptasi yang unik. Penelitian ini bertujuan untuk mengidentifikasi filogenetik isolat bakteri dari tambak garam Osowilangun menggunakan pendekatan molekuler berbasis gen 16S rRNA. Proses isolasi dilakukan pada sampel air tambak menggunakan media Nutrient Agar (NA) dan Zobell Marine Broth (ZMB) yang dimodifikasi dengan 10% NaCl. Dua isolat terpilih, TGA-2 dan TGA-7, berhasil dimurnikan dan diregenerasi. Hasil pewarnaan Gram menunjukkan bahwa kedua isolat merupakan bakteri Gram positif berbentuk kokus. Kurva pertumbuhan menunjukkan fase stasioner rata–rata pada jam ke-21 hingga jam ke-80. Ekstraksi DNA menghasilkan konsentrasi tinggi dengan kemurnian baik, dan amplifikasi gen 16S rRNA menghasilkan pita DNA tunggal sepanjang ±1500 bp. Hasil sekuensing dianalisis menggunakan metode Neighbour-Joining pada perangkat lunak MEGA 12. Analisis filogenetik menunjukkan bahwa isolat TGA-2 memiliki kemiripan 98,73% dengan Staphylococcus ureilyticus strain HBU7259, sedangkan isolat TGA-7 menunjukkan kemiripan 99,02% dengan Staphylococcus cohnii strain Tr15B. Kedua isolat tersebut termasuk dalam genus Staphylococcus koagulase-negatif, yang diidentifikasikan sebagai halotoleran bukan halofilik, karena tetap mampu tumbuh tanpa garam. Hasil penelitian ini memberikan data identifikasi molekuler dan hubungan filogenetik isolat bakteri dari tambak garam Osowilangun Surabaya, yang dapat dikembangkan lebih lanjut pada studi bioaktivitas berikutnya.
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Hypersaline environments such as the Osowilangun salt pond in Surabaya represent extreme ecosystems that potentially contain a halophilic microorganism, including bacteria with unique adaptive capabilities. This study aimed to identify the phylogenetic relationships of bacterial isolates obtained from the Osowilangun salt pond using a molecular approach based on the 16S rRNA gene. Isolation was performed on pond water samples using Nutrient Agar (NA) and Zobell Marine Broth (ZMB) modified with 10% NaCl. Two selected isolates, TGA-2 and TGA-7, were successfully purified and regenerated. Gram staining indicated that both isolates were Gram-positive cocci. Growth curve analysis showed that the stationary phase occurred, on average, between the 21st and 80th hours. DNA extraction yielded high concentrations with good purity, and amplification of the 16S rRNA gene produced a single DNA band of approximately 1500 bp. Sequencing results were analyzed using the Neighbor-Joining method in the MEGA 12 software. Phylogenetic analysis revealed that isolate TGA-2 shared 98.73% similarity with Staphylococcus ureilyticus strain HBU7259, whereas isolate TGA-7 showed 99.02% similarity with Staphylococcus cohnii strain Tr15B. Both isolates belong to the coagulase-negative Staphylococcus group and were identified as halotolerant rather than truly halophilic, as they were able to grow even in the absence of salt. This study provides molecular identification and phylogenetic analysis of bacterial isolates from the Osowilangun salt ponds, Surabaya, which may serve as a basis for futher investigation in bioactivity studies.

Item Type:	Thesis (Other)
Uncontrolled Keywords:	Tambak garam, Isolasi, Halotoleran, Ekstraksi DNA, Gen 16S rRNA, Filogenetik, Salt pond, Isolate, Halotolerant, DNA extraction, 16S rRNA gene, Phylogenetics
Subjects:	Q Science > QD Chemistry > QD251.2 Chemistry, Organic. Biochemistry
Divisions:	Faculty of Science and Data Analytics (SCIENTICS) > Chemistry > 47201-(S1) Undergraduate Thesis
Depositing User:	Muhammad Alvan Thariq Risqullah
Date Deposited:	04 Feb 2026 01:53
Last Modified:	04 Feb 2026 01:53
URI:	http://repository.its.ac.id/id/eprint/131994

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