Identifikasi dan Karakterisasi Isolat Bifidobacterium breve BS2-PS2 dari Air Susu Ibu Sebagai Kandidat Probiotik

Rombot, Angle Berliani (2023) Identifikasi dan Karakterisasi Isolat Bifidobacterium breve BS2-PS2 dari Air Susu Ibu Sebagai Kandidat Probiotik. Bachelor thesis, Universitas Pelita Harapan.

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Abstract

Bifidobacterium sp. merupakan bakteri yang umum dimanfaatkan sebagai probiotik. Namun, penelitian terkait identifikasi dan karakterisasi Bifidobacterium sp. dari ASI belum banyak dilakukan di Indonesia. Pada penelitian ini, dilakukan identifikasi dan karakterisasi awal isolat Bifidobacterium sp. BS2-PS2 sebagai kandidat probiotik. Penelitian dilakukan dengan tahap sebagai berikut: identifikasi berdasarkan morfologi koloni dengan pengamatan koloni, morfologi sel berdasarkan pewarnaan Gram, pewarnaan endospora, karakterisasi biokima dengan uji katalase, identifikasi molekuler dengan 16S rRNA sequencing, kemudian dilakukan juga karakterisasi awal sifat probiotik meliputi uji ketahanan: terhadap phenol konsentrasi 0,2% dan 0,5%; NaCl konsentrasi 1,5%, 2,5%, dan 3,5%; garam empedu konsentrasi 0,3% dan 0,5%; dan simulated gastric juice. Hasil analisis: morfologi koloni BS2-PS2 berupa bentuk bulat, warna putih, margin entire, dan konveks; pewarnaan Gram positif; morfologi sel berbentuk batang dengan percabangan yang bervariasi menyerupai bentuk V dan Y (bifid); pewarnaan endospora negatif; uji katalase negatif; identifikasi molekuler dengan 16S rRNA sequencing menunjukkan isolat berupa spesies Bifidobacterium breve (query cover: 99%; tingkat identitas: 98%); karakteristik sifat probiotik: tidak memiliki ketahanan terhadap phenol pada konsentrasi 0,2% dan 0,5%; memiliki ketahanan NaCl pada konsentrsasi 1,5%, tidak tahan NaCl pada konsentrasi 2,5% dan 3,5%; tahan pada garam empedu dengan konsentrasi 0,3% dan 0,5 %; dan tahan terhadap simulated gastric juice. Dari semua hasil percobaan tersebut, dapat disimpulkan bahwa B. breve BS2-PS2 telah memenuhi kriteria awal sebagai kandidat probiotik/Bifidobacterium sp. are commonly used as probiotics. However, research related to the identification and characterization of Bifidobacterium spp. from breast milk has not been widely carried out in Indonesia. In this research, identification and initial characterization of Bifidobacterium sp. BS2-PS2 as probiotic candidate was carried out. The research was carried out in the following stages: identification based on colony morphology by observing colonies, cell morphology by Gram staining, endospore staining, biochemical characterization by catalase test, molocular identification by 16S rRNA sequencing, then some initial characterization of probiotic properties were carried out including resistance tests to: phenol concentrations of 0.2% and 0.5%; NaCl concentration of 1.5%. 2.5% and 3.5%; bile salt concentration of 0.3% and 0.5%; and simulated gastric juice. Result of analysis: colony of BS2-PS2 have a morphology with a round shape, white color, entire margin, and convex elevation; Gram positive bacteria; rod-shape cell morphology with branches remebling a “V and Y” shape (bifid); non endospore-forming; negative catalase test; molecular identification by 16S rRNA sequencing showed the isolate to be a species of Bifidobacterium breve (query cover: 99%; percent identity: 98%); characteristics of probiotic properties: not resistance to phenol at concentrations of 0.2% and 0.5%; resistance to NaCl at a concentration of 1.5%, not resistant to NaCl at concentrations of 2.5% and 3.5%; resistant to bile salts with concentration of 0.3% and 0.5%; and resistant to simulated gastric juice. From all the experimental result above, it can be concluded that B. breve BS2-PS2 has fulfilled the initial criteria as a probiotic candidate.

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