Isolasi dan identifikasi bakteri Bifidobacterium sp. dari air susu ibu (ASI) = Isolation and identification of Bifidobacterium sp. from human breast milk

Rachmah, Athiyyarizka Farbila (2020) Isolasi dan identifikasi bakteri Bifidobacterium sp. dari air susu ibu (ASI) = Isolation and identification of Bifidobacterium sp. from human breast milk. Bachelor thesis, Universitas Pelita Harapan.

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Abstract

ASI merupakan sumber nutrisi satu-satunya yang penting untuk bayi dan mengandung mikrobiota probiotik penting untuk perkembangan kesehatan bayi. Salah satu anggota mikrobiota yang diisolasi dari ASI adalah Bifidobacteria. Bifidobacteria berperan penting pada pencernaan bayi karena mampu memetabolisme HMO (human milk oligosaccharide) dari ASI. Penelitian ini bertujuan untuk mengisolasi, mengidentifikasi, serta mengkarakterisasi bakteri anaerobik obligat Bifidobacterium sp. dari ASI yang sejauh ini belum dilakukan di Indonesia. Pada penelitian ini dua sampel ASI kolostrum (SA-1, SA-2) dari hari ke 1-2 digunakan untuk isolasi Bifidobacteria. Metode yang dilakukan pada penelitian ini antara lain adalah; menumbuhkan bakteri dari sampel ASI pada media MRS yang disuplementasikan L-csyteine dan antibiotik Mupirocin atau MRSC-MUP1, media MRSC-MUP yang ditambahkan susu sapi (MRSC-MUP2), serta media TPY (trypticase phytone yeast extract) serta identifikasi morfologi, dan karakterisasi isolat (uji aktivitas enzim katalase dan uji motilitas agar). Potensi isolat dari sampel ASI sebagai probiotik juga dievaluasi melalui uji ketahanan pH; uji ketahanan garam empedu; dan uji suseptibilitas terhadap berbagai antibiotik. Identifikasi molekuler juga dilakukan dan dimulai dengan ekstraksi, PCR, elektroforesis, dan sequencing 16S-rRNA. Berdasarkan hasil yang diperoleh, jenis media terbaik untuk pertumbuhan bakteri dari sampel ASI adalah media TPY. Rata-rata CFU/ml dari sampel ASI SA-1 dan SA-2 masing-masing sebanyak 2 x 10^5 dan 7 x 10^3. Isolat SU-KC1 yang diperoleh dari sampel ASI SA-1 memiliki karakterisik gram-positif, acid-fast negatif, non-motil, dan katalase positif. Dari hasil evaluasi probiotik, isolat SU-KC1 berpotensi sebagai probiotik. Berdasarkan hasil identifikasi molekuler dengan primer spesifik Bif164 dan Bif662 dan hasil 16S-rRNA, isolat SU-KC1 diduga merupakan spesies Bifidobacterium sp. terbaru yang belum diidentifikasi, namun memiliki kesamaan dengan B. actinocoloniiforme strain DSM 22766, B. actinocoloniiforme strain JCM 18048 dan B. xylocopae strain XV2 yang memiliki kekerabatan dengan golongan Bifidobacteria katalase positif yaitu B. adolescentis dan B. indicum. Selain itu, penelitian lebih lanjut perlu dilakukan untuk mendukung potensi isolat sebagai probiotik yang aman digunakan. / Breast milk is an important source of nutrition and probiotics for infants during their early development. Bifidobacterium sp. is one of the most dominant microbes found in breast milk and is considered the key member of infant’s gut microbiota through its ability to metabolize HMO (human milk oligosaccharides) from breast milk. Isolation of Bifidobacterium sp. from breast milk samples in Indonesia has not been reported. This research aims to isolate, identify and characterize Bifidobacterium sp. from breast milk and evaluate its potential as probiotic. Two colostrum (SA-1, SA-2) from day 1-2 breast milk samples were obtained and used for Bifidobacteria isolation. Different types of Bifidobacteria selective media such as MRS agar supplemented with L-cysteine and Mupirocin antibiotic (MRSC-MUP1), MRSC-MUP media supplemented with bovine milk (MRSC-MUP2), and TPY media (trypticase phytone yeast extract) were used in this research. The probiotic potential from breast milk samples were also evaluated through acidity and bile salt tolerance and antibiotic susceptibility test. Molecular identification is done through DNA extraction, PCR analysis, gel visualization, and 16S-rRNA sequencing. TPY media is found to be the most suitable media to grow Bifidobacteria from the breast milk samples obtained. The average CFU/ml from each breast milk samples (SA-1 and SA-2) are 2 x 10^5 and 7 x 10^3 respectively. The isolate SU-KC1 obtained from breast milk sample SA-1, is gram positive, acid-fast negative, non-motile, and catalase positive. Specific primer Bif164 and Bif662 is used in PCR analysis. 16S-rRNA sequencing results indicates that isolate SU-KC1 belong to a novel species within the genus Bifidobacterium sp. and shows similar identity with B. actinocoloniiforme strain DSM 22766, B. actinocoloniiforme strain JCM 18048 and B. xylocopae strain XV2, which were affiliated to members of catalase-positive Bifidobacteria; B. adolescentis and B. indicum. Results also indicates that isolate SU-KC1 have the potential to be used as probiotic, however further research is required to support this claim.

Item Type: Thesis (Bachelor)
Creators:
CreatorsNIMEmail
Rachmah, Athiyyarizka FarbilaNIM00000021569farbilarizka@gmail.com
Contributors:
ContributionContributorsNIDN/NIDKEmail
Thesis advisorTan, Tjie JanNIDN0318104902tjiejan.tan@gmail.com
Thesis advisorSugata, MarceliaNIDN0321118902marcelia.sugata@uph.edu
Additional Information: SK 113-16 RAC i
Uncontrolled Keywords: isolasi; identifikasi; Bifidobacteria; air susu ibu (ASI)
Subjects: Q Science > QH Natural history > QH301 Biology
Divisions: University Subject > Current > Faculty/School - UPH Karawaci > Faculty of Science and Technology > Biology
Current > Faculty/School - UPH Karawaci > Faculty of Science and Technology > Biology
Depositing User: Users 3741 not found.
Date Deposited: 23 Sep 2020 03:23
Last Modified: 07 Oct 2021 03:01
URI: http://repository.uph.edu/id/eprint/11061

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